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1.
Medicine (Baltimore) ; 100(40): e27433, 2021 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-34622857

RESUMO

ABSTRACT: There are approximately 2 billion HBV-infected individuals worldwide, and approximately 1.87% to 7% of these individuals are copositive for HBsAg and HBsAb.Our study detected hepatitis B virus pgRNA (HBV RNA) levels in HBsAg and HBsAb copositive patients and then analyzed the correlation with HBV DNA, HBsAg, ALT, and AST levels. A total of 149 HBsAg and HBsAb copositive patients were identified from 66,617 outpatients.HBV RNA, HBV DNA, HBsAg, ALT, and AST serum levels were significantly different in different natural phases of HBV infection (immune tolerance phase, immune clearance phase, low replication phase, and reactivation phase) with statistical significance (P < .01). HBV RNA levels were positively correlated with HBV DNA, HBsAg, ALT, and AST levels. HBV RNA and HBV DNA levels were significantly increased in the HBeAg-positive group (66 patients) compared with the HBeAg-negative group (83 patients) (P < .01). In the HBeAg-positive group, HBV RNA levels were positively correlated with HBV DNA and HBsAg levels. In the HBeAg-negative group, HBV RNA levels were positively correlated with HBV DNA. Serum HBV RNA levels were positively correlated with HBV DNA, HBsAg, ALT, and AST levels.HBV RNA could be used as a virological indicator for antiviral therapy in HBsAg and HBsAb copositive hepatitis B patients.


Assuntos
DNA Viral/sangue , Vírus da Hepatite B/genética , Hepatite B Crônica/virologia , Adulto , Progressão da Doença , Feminino , Genótipo , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Hepatite B Crônica/sangue , Humanos , Masculino , Pessoa de Meia-Idade
2.
Pan Afr Med J ; 39: 175, 2021.
Artigo em Francês | MEDLINE | ID: mdl-34584601

RESUMO

Introduction: viral infection caused by hepatitis B virus is the most frequent transfusion-transmitted viral infection. Although the search for hepatitis B surface antigen (HBsAg) in blood banks has significantly reduced the risk for transfusion-transmitted virus infection, there is still a residual transfusion risk of transmission from donors with occult hepatitis B. Blood bags containing aHBc with or without aHBs and viral DNA can cause infections and represent a threat to transfusion safety when aHBc levels are undetectable. The purpose of this study is to determine the residual risk for transfusion-transmitted hepatitis B virus at the Central Hospital of Yaoundé (CHY) as well as at the St Martin de Porres's Catholic Hospital (SMPCH) in Yaoundé, Cameroon. Methods: we conducted a cross-sectional study among blood donors at the Central Hospital of Yaoundé (CHY) and the St Martin de Porres's Catholic Hospital. In these subjects the search for aHBc and/or the aHBs was conducted by immunochromatography. HBV DNA test was performed on blood samples tested positive for aHBc and/or aHBs by Polymerase Chain Reaction (PCR) technique using specific primers. Results: out of a total of 193 blood donors negative for HIV, HBV (HBsAg), HCV serological markers and treponema infections, the overall seroprevalence of aHBc and/or aHBs was 9,84% (19/193). Out of a total of 19 potentially infected donors, HBV DNA was detected in 03 individuals, including 02 aHBc carriers and 01 carrier of both aHBc and aHBs, reflecting a prevalence of occult hepatitis B of 15,79% (3/19) [IC 95% =3,38%-39,58%] and a residual risk for transfusion-transmitted hepatitis B virus of 1,55% (3/193) [IC 95% =0,32%-4,48%]. Conclusion: this study shows that the residual risk for transfusion-transmitted hepatitis B virus is low. However, it is recommended to screan blood donors for aHBc and/or aHBs.


Assuntos
Doadores de Sangue , Seleção do Doador/métodos , Antígenos de Superfície da Hepatite B/sangue , Hepatite B/transmissão , Adolescente , Adulto , Segurança do Sangue/métodos , Transfusão de Sangue/normas , Camarões , Estudos Transversais , DNA Viral/sangue , Feminino , Hepatite B/sangue , Hepatite B/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Medição de Risco , Estudos Soroepidemiológicos , Adulto Jovem
3.
Arch Virol ; 166(11): 3075-3084, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34468889

RESUMO

Occult hepatitis B virus (OBI) infection is defined by the presence of viral DNA in the liver and/or serum in absence of hepatitis B surface antigen (HBsAg). While multiple studies have identified mutations that are associated with OBI, only a small portion of these mutations have been functionally characterized in vitro. Using complementary in silico approaches, the effects of OBI-associated amino acid mutations on HBV protein function in HBV/HIV-positive ART-naïve South Africans were evaluated. Two OBI-associated mutations in the PreS1 region, one in the PreS2 region, and seven in the surface region of subgenotype A1 sequences were identified as deleterious. In subgenotype A2 sequences, 11 OBI-associated mutations in the PreS1 region, seven in the PreS2 region, and 31 in the surface region were identified as deleterious. In the polymerase region, 14 OBI-associated mutations in subgenotype A1 and 71 OBI-associated mutations in subgenotype A2 were identified as deleterious. This study utilized in silico approaches to characterize the likely impact of OBI-associated mutations on viral function, thereby identifying and prioritizing candidates and reducing the significant cost associated with functional studies that are essential for mechanistic studies of the OBI phenotype.


Assuntos
Vírus da Hepatite B/genética , Hepatite B/virologia , Mutação , Simulação por Computador , DNA Viral/sangue , Genótipo , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/química , Vírus da Hepatite B/patogenicidade , Humanos , África do Sul
4.
Transfus Med ; 31(5): 371-376, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34390068

RESUMO

BACKGROUND: Pathogen reduction technologies (PRT) based on nucleic-acid damaging chemicals and/or irradiation are increasingly being used to increase safety of blood components against emerging pathogens, such as convalescent plasma in the ongoing COVID-19 pandemic. Current methods for PRT validation are limited by the resources available to the blood component manufacturer, and quality control rely over pathogen spiking and hence invariably require sacrifice of the tested blood units: quantitative real-time PCR is the current pathogen detection method but, due to the high likelihood of detecting nonviable fragments, requires downstream pathogen culture. We propose here a new molecular validation of PRT based on the highly prevalent human symbiont torquetenovirus (TTV) and rolling circle amplification (RCA). MATERIALS AND METHODS: Serial apheresis plasma donations were tested for TTV before and after inactivation with Intercept® PRT using real-time quantitative PCR (conventional validation), RCA followed by real-time PCR (our validation), and reverse PCR (for cross-validation). RESULTS: While only 20% of inactivated units showed significant decrease in TTV viral load using real-time qPCR, all donations tested with RCA followed by real-time PCR showed TTV reductions. As further validation, 2 units were additionally tested with reverse PCR, which confirmed absence of entire circular genomes. DISCUSSION: We have described and validated a conservative and easy-to-setup protocol for molecular validation of PRT based on RCA and real-time PCR for TTV.


Assuntos
DNA Viral , Reação em Cadeia da Polimerase em Tempo Real , Torque teno virus , Inativação de Vírus , COVID-19/sangue , COVID-19/genética , DNA Viral/sangue , DNA Viral/genética , Humanos , SARS-CoV-2/genética , SARS-CoV-2/metabolismo , Torque teno virus/genética , Torque teno virus/metabolismo
5.
Sci Rep ; 11(1): 16982, 2021 08 20.
Artigo em Inglês | MEDLINE | ID: mdl-34417517

RESUMO

HBV infection is recognized as a serious global health problem, and hepatitis B virus infection is a complicated chronic disease leading to liver cirrhosis (LC) and hepatocellular carcinoma (HCC). New biochemical serum markers could be used to advance the diagnosis and prognosis of HBV-associated liver diseases during the progression of chronic hepatitis B into cirrhosis and HCC. We determined whether the 4210 Da and 1866 Da polypeptides are serum metabolite biomarkers of hepatopathy with hepatitis B virus. A total of 570 subjects were divided into five groups: healthy controls, those with natural clearance, and patients with CHB, LC, and HCC. The 1866 Da and 4210 Da polypeptides were measured by Clin-ToF II MALDI-TOF-MS. There were significant differences in 4210 Da and 1866 Da levels among the five groups (P < 0.001). For the differential diagnosis of CHB from normal liver, the areas under the receiver operating characteristic (ROC) curve of 4210 Da and 1866 Da and their combination via logistic regression were 0.961, 0.849 and 0.967. For the differential diagnosis of LC from CHB, the areas under the ROC curve were 0.695, 0.841 and 0.826. For the differential diagnosis of HCC from CHB, the areas under the ROC curve were 0.744, 0.710 and 0.761, respectively. For the differential diagnosis of HCC from LC, the areas under the ROC curve of 4210 Da and 1866 Da were 0.580 and 0.654. The positive rate of 1866 Da was 45.5% and 69.0% in AFP-negative HCC patients and that of 4210 Da was 60.6% 58.6% in AFP-negative HCC patients of the study HCC vs. CHB and HCC vs. LC. The 4210 Da and 1866 Da polypeptide levels were positively correlated with HBV DNA levels (P < 0.001, r = 0.269; P < 0.001, r = 0.285). The 4210 Da and 1866 Da polypeptides had good diagnostic value for the occurrence and progression of HBV-related chronic hepatitis, liver cirrhosis and hepatocellular carcinoma and could serve to accurately guide treatment management and predict clinical outcomes.


Assuntos
Progressão da Doença , Vírus da Hepatite B/fisiologia , Hepatopatias/patologia , Hepatopatias/virologia , Peptídeos/metabolismo , Biomarcadores/sangue , Biomarcadores/metabolismo , Carcinoma Hepatocelular/sangue , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/virologia , DNA Viral/sangue , Feminino , Hepatite B Crônica/sangue , Hepatite B Crônica/diagnóstico , Hepatite B Crônica/patologia , Hepatite B Crônica/virologia , Humanos , Hepatopatias/sangue , Hepatopatias/diagnóstico , Neoplasias Hepáticas/sangue , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Peso Molecular , Peptídeos/sangue , Curva ROC , alfa-Fetoproteínas/metabolismo
6.
PLoS One ; 16(8): e0254136, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34383785

RESUMO

Human cytomegalovirus (HCMV) is a highly prevalent herpes virus which persists as a latent infection and has been detected in several different tumor types. HCMV disease is rare but may occur in high-risk settings, often manifesting as a pulmonary infection. To date HCMV has not been investigated in malignant pleural mesothelioma (MPM). In a consecutive case series of 144 MPM patients we evaluated two biomarkers of HCMV: IgG serostatus (defined as positive and negative) and DNAemia (>100 copies/mL of cell free HCMV DNA in serum). Approximately half of the MPM patient population was HCMV IgG seropositive (51%). HCMV DNAemia was highly prevalent (79%) in MPM and independent of IgG serostatus. DNAemia levels consistent with high level current infection (>1000 copies/mL serum) were present in 41% of patients. Neither IgG serostatus nor DNAemia were associated with patient survival. In tissues, we observed that HCMV DNA was present in 48% of tumors (n = 40) and only 29% of normal pleural tissue obtained from individuals without malignancy (n = 21). Our results suggest nearly half of MPM patients have a high level current HCMV infection at the time of treatment and that pleural tissue may be a reservoir for latent HCMV infection. These findings warrant further investigation to determine the full spectrum of pulmonary infections in MPM patients, and whether treatment for high level current HCMV infection may improve patient outcomes.


Assuntos
Anticorpos Antivirais/sangue , Citomegalovirus/metabolismo , DNA Viral/sangue , Imunoglobulina G/sangue , Mesotelioma Maligno , Neoplasias Pleurais , Pneumonia Viral , Idoso , Infecções por Citomegalovirus/sangue , Infecções por Citomegalovirus/mortalidade , Intervalo Livre de Doença , Feminino , Humanos , Masculino , Mesotelioma Maligno/sangue , Mesotelioma Maligno/mortalidade , Mesotelioma Maligno/virologia , Pessoa de Meia-Idade , Neoplasias Pleurais/sangue , Neoplasias Pleurais/mortalidade , Neoplasias Pleurais/virologia , Pneumonia Viral/sangue , Pneumonia Viral/mortalidade , Pneumonia Viral/virologia , Estudos Retrospectivos , Taxa de Sobrevida
7.
Nat Commun ; 12(1): 4181, 2021 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-34234136

RESUMO

Nucleobase and nucleoside analogs (NNA) are widely used as anti-viral and anti-cancer agents, and NNA phosphorylation is essential for the activity of this class of drugs. Recently, diphosphatase NUDT15 was linked to thiopurine metabolism with NUDT15 polymorphism associated with drug toxicity in patients. Profiling NNA drugs, we identify acyclovir (ACV) and ganciclovir (GCV) as two new NNAs metabolized by NUDT15. NUDT15 hydrolyzes ACV and GCV triphosphate metabolites, reducing their effects against cytomegalovirus (CMV) in vitro. Loss of NUDT15 potentiates cytotoxicity of ACV and GCV in host cells. In hematopoietic stem cell transplant patients, the risk of CMV viremia following ACV prophylaxis is associated with NUDT15 genotype (P = 0.015). Donor NUDT15 deficiency is linked to graft failure in patients receiving CMV-seropositive stem cells (P = 0.047). In conclusion, NUDT15 is an important metabolizing enzyme for ACV and GCV, and NUDT15 variation contributes to inter-patient variability in their therapeutic effects.


Assuntos
Aciclovir/farmacologia , Antivirais/farmacologia , Infecções por Citomegalovirus/prevenção & controle , Ganciclovir/análogos & derivados , Pirofosfatases/genética , Aciclovir/uso terapêutico , Adolescente , Adulto , Idoso , Animais , Antibioticoprofilaxia , Antivirais/uso terapêutico , Variação Biológica da População/genética , Linhagem Celular , Criança , Pré-Escolar , Cristalografia por Raios X , Citomegalovirus/efeitos dos fármacos , Citomegalovirus/genética , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/diagnóstico , Infecções por Citomegalovirus/etiologia , Infecções por Citomegalovirus/virologia , DNA Viral/sangue , DNA Viral/isolamento & purificação , Modelos Animais de Doenças , Farmacorresistência Viral , Feminino , Ganciclovir/farmacologia , Ganciclovir/uso terapêutico , Transplante de Células-Tronco Hematopoéticas/efeitos adversos , Interações entre Hospedeiro e Microrganismos/genética , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Muromegalovirus/isolamento & purificação , Muromegalovirus/patogenicidade , Variantes Farmacogenômicos , Polimorfismo de Nucleotídeo Único , Pirofosfatases/metabolismo , Pirofosfatases/ultraestrutura , Resultado do Tratamento , Adulto Jovem
8.
BMC Infect Dis ; 21(1): 714, 2021 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-34330225

RESUMO

BACKGROUND: Nucleic acid amplification testing (NAT) for blood screening has been previously performed in some countries to determine NAT yields. The current study sought to explore the non-discriminating reactive NAT yields using individual-NAT (ID-NAT) and characteristics of HBV NAT yields through a 10-year retrospective analysis in Zhejiang, China. METHODS: Blood donations were analyzed using individual-NAT mode by the transcription-mediated amplification (TMA) method. Supplementary HBV serological tests were performed using chemiluminescent immunoassay, and HBV viral load assay was performed by real-time polymerase chain reaction. Follow-up studies were performed in partial donors with low HBV viral loads. RESULTS: Non-discriminating reactive NAT yields and HBV NAT yields varied in different years. The yields ranged from 853.73 per million to 2018.68 per million and 624.60 per million to 1669.50 per million, respectively. In the 476 NAT yields, 19 were probable window periods (WP), 33 probable occult hepatitis B virus infections (OBIs), 409 were confirmed OBIs and 15 were chronic HBV infections. ID-NAT results were categorized in four groups, and the findings showed that the levels of HBV DNA viral loads were different in the four different groups (χ2 = 275.02, p < 0.01). HBV viral load distribution was significantly different between anti-HBs positive and anti-HBc positive samples (χ2 = 49.429, p < 0.01). Notably, only 42.03% donors were NAT repeated positive in the 138 repeat donors' follow up tests. CONCLUSION: NAT screening of blood donations can reduce the risk of transfusion-transmitted HBV infections. Positive proportions of anti-HBs and anti-HBc are correlated with the HBV viral load level. However, low level of viral load donors pose risks in HBV NAT assays, and show fluctuating state for HBV viral load and leads to non-repeated NAT results during follow up studies.


Assuntos
Doadores de Sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B/prevenção & controle , Técnicas de Amplificação de Ácido Nucleico , Reação Transfusional/prevenção & controle , China , DNA Viral/sangue , Hepatite B/diagnóstico , Anticorpos Anti-Hepatite B/sangue , Vírus da Hepatite B/genética , Humanos , Programas de Rastreamento , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Testes Sorológicos , Carga Viral
9.
BMC Pregnancy Childbirth ; 21(1): 513, 2021 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-34273944

RESUMO

BACKGROUND: We aimed to investigate whether maternal chronic hepatitis B virus (HBV) infection affects preterm birth (PTB) in pregnant women. METHODS: We retrospectively analyzed HBV-infected and non-infected pregnant women attending antenatal care at Fujian Maternity and Child Health Hospital, Fuzhou, China between January 1, 2016 to December 31, 2018. Participants were divided into HBV infection (n = 1302) and control (n = 12,813) groups. We compared baseline data, pregnancy and perinatal complications, and preterm delivery outcomes between groups. Performed multiple logistics regression analysis to adjust for confounding factors. Finally, we compared early PTB outcome between different HBV DNA level groups. RESULTS: The incidence of preterm birth (gestation less than 37 weeks) was similar between the groups, early preterm birth (gestation less than 34 weeks) were significantly more among the HBV infection group than among the controls (1.6% VS. 0.8%; P = 0.003). After adjusting for confounding factors through logistics regression, HBV infection was found to be an independent early PTB risk factor gestation (adjusted odds ratio 1.770; 95% confidence interval [1.046-2.997]). The incidence of early PTB in < 500 group, 500 ~ 2.0 × 10e5 group and > 2.0 × 10e5 group was not statistically significant (P = 0.417). CONCLUSION: HBV infection is an independent risk factor for early PTB, and the risk did not seem to be influenced by the levels of HBV DNA. Comprehensive programs focusing on pregnant women with HBV infection would reduce the incidence of adverse outcomes.


Assuntos
Hepatite B Crônica/epidemiologia , Hepatite B/epidemiologia , Complicações Infecciosas na Gravidez/epidemiologia , Nascimento Prematuro/epidemiologia , Adulto , China/epidemiologia , Estudos de Coortes , DNA Viral/sangue , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Fatores de Risco , Carga Viral
10.
BMC Infect Dis ; 21(1): 592, 2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34157979

RESUMO

BACKGROUND: Factors predicting peripheral blood total HIV-1 DNA size in chronically infected patients with successfully suppressed viremia remain unclear. Prognostic power of such factors are of clinical significance for making clinical decisions. METHODS: Two sets of study populations were included: 490 China AIDS Clinical Trial (CACT) participants (Training cohort, followed up for 144 to 288 weeks) and 117 outpatients from Peking Union Medical College Hospital (PUMCH) (Validation cohort, followed up for more than 96 weeks). All patients were chronically HIV-1-infected and achieved successful HIV-1 plasma RNA suppression within week 48. Total HIV-1 DNA in blood at baseline, 12, 24, 48, 96, 144 and 288 weeks after combined antiretroviral therapy (cART) initiation were quantified. Generalized estimating equations and logistic regression methods were used to derive and validate a predictive model of total HIV-1 DNA after 96 weeks of cART. RESULTS: The total HIV-1 DNA rapidly decreased from baseline [median = 3.00 log10 copies/106 peripheral blood mononuclear cells (PBMCs)] to week 24 (median = 2.55 log10 copies/106 PBMCs), and leveled off afterwards. Of the 490 patients who had successful HIV-1 plasma RNA suppression by 96 w post-cART, 92 (18.8%) had a low total HIV-1 DNA count (< 100 copies/106 PBMCs) at week 96. In the predictive model, lower baseline total HIV-1 DNA [risk ratio (RR) = 0.08, per 1 log10 copies/106 PBMCs, P < 0.001] and higher baseline CD4+ T cell count (RR = 1.72, per 100 cells/µL, P < 0.001) were significantly associated with a low total HIV-1 DNA count at week 96. In an independent cohort of 117 patients, this model achieved a sensitivity of 75.00% and specificity of 69.52%. CONCLUSIONS: Baseline total HIV-1 DNA and CD4+ T cell count are two independent predictors of total HIV-1 DNA after treatment. The derived model based on these two baseline factors provides a useful prognostic tool in predicting HIV-1 DNA reservoir control during cART.


Assuntos
DNA Viral/sangue , HIV-1 , Leucócitos Mononucleares/virologia , Modelos Estatísticos , Carga Viral , Adulto , Antirretrovirais/uso terapêutico , Contagem de Linfócito CD4 , China/epidemiologia , Estudos de Coortes , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Humanos , Estudos Longitudinais , Masculino , Sensibilidade e Especificidade , Viremia/tratamento farmacológico
11.
Pan Afr Med J ; 38: 294, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34178213

RESUMO

Introduction: quantification of hepatitis B virus DNA, a key element in the management of chronic hepatitis B, allows a more direct and reliable measurement of viral replication and monitoring of the virological response to therapy. Polymerase chain reaction (PCR) platforms performing this quantification and adaptable to intermediate laboratories have been developed. Thus, this study was conducted to evaluate the on-site performance of the AMPLIX® hepatitis B virus (HBV) real-time PCR technique in comparison with the COBAS AmpliPrep™ technique. Methods: performance of the AMPLIX® HBV real-time PCR technique was evaluated with repeatability and intermediate precision (reproducibility) determined. The comparison with COBAS Taqman was performed by testing, in parallel, 42 plasma samples. The statistical analysis using Meth Val® software was focused on correlation and concordance determination. Results: AMPLIX® real-time PCR assay showed good reproducibility for the low (CV=6.65%) and high (CV=3.15%) control levels but also good repeatability for both the low (CV=2.12%) and high (CV=1.60%) concentration levels. Accuracy obtained in our study were less than acceptability limit fixed to 5%. Viral load measurements between Amplix and COBAS Taqman correlated strongly with a correlation coefficient of 0.97%. Concordance analysis gave an average of the differences of 0.54 log IU/L between the viral load measurements of the 2 techniques. Conclusion: based on these results, the Amplix real-time PCR platform for the quantification of HBV DNA can be considered as a reliable system for the monitoring of chronic hepatitis B and also a system adapted to intermediate laboratories.


Assuntos
Vírus da Hepatite B/isolamento & purificação , Hepatite B Crônica/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA Viral/sangue , Vírus da Hepatite B/genética , Hepatite B Crônica/virologia , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes , Senegal , Carga Viral
12.
Nat Commun ; 12(1): 3727, 2021 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-34140517

RESUMO

Clonal expansion of HIV-infected cells contributes to the long-term persistence of the HIV reservoir in ART-suppressed individuals. However, the contribution from cell clones that harbor inducible proviruses to plasma viremia is poorly understood. Here, we describe a single-cell approach to simultaneously sequence the TCR, integration sites and proviral genomes from translation-competent reservoir cells, called STIP-Seq. By applying this approach to blood samples from eight participants, we show that the translation-competent reservoir mainly consists of proviruses with short deletions at the 5'-end of the genome, often involving the major splice donor site. TCR and integration site sequencing reveal that cell clones with predicted pathogen-specificity can harbor inducible proviruses integrated into cancer-related genes. Furthermore, we find several matches between proviruses retrieved with STIP-Seq and plasma viruses obtained during ART and upon treatment interruption, suggesting that STIP-Seq can capture clones that are responsible for low-level viremia or viral rebound.


Assuntos
Antirretrovirais/uso terapêutico , Infecções por HIV/sangue , Infecções por HIV/tratamento farmacológico , HIV-1/metabolismo , Provírus/genética , Análise de Célula Única/métodos , Viremia/virologia , Linfócitos T CD4-Positivos/virologia , DNA Viral/sangue , Infecções por HIV/virologia , HIV-1/genética , HIV-1/patogenicidade , Humanos , Ionomicina/farmacologia , Masculino , Pessoa de Meia-Idade , Filogenia , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Deleção de Sequência , Carga Viral/genética
13.
Nat Commun ; 12(1): 3922, 2021 06 29.
Artigo em Inglês | MEDLINE | ID: mdl-34188039

RESUMO

Non-invasive biomarkers that predict HIV remission after antiretroviral therapy (ART) interruption are urgently needed. Such biomarkers can improve the safety of analytic treatment interruption (ATI) and provide mechanistic insights into the host pathways involved in post-ART HIV control. Here we report plasma glycomic and metabolic signatures of time-to-viral-rebound and probability-of-viral-remission using samples from two independent cohorts. These samples include a large number of post-treatment controllers, a rare population demonstrating sustained virologic suppression after ART-cessation. These signatures remain significant after adjusting for key demographic and clinical confounders. We also report mechanistic links between some of these biomarkers and HIV latency reactivation and/or myeloid inflammation in vitro. Finally, machine learning algorithms, based on selected sets of these biomarkers, predict time-to-viral-rebound with 74% capacity and probability-of-viral-remission with 97.5% capacity. In summary, we report non-invasive plasma biomarkers, with potential functional significance, that predict both the duration and probability of HIV remission after treatment interruption.


Assuntos
Biomarcadores/sangue , Infecções por HIV/sangue , Suspensão de Tratamento , Adulto , Antirretrovirais/administração & dosagem , Estudos de Coortes , DNA Viral/sangue , Feminino , Glicômica , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , Humanos , Inflamação , Macrófagos/imunologia , Masculino , Metabolômica , Pessoa de Meia-Idade , Modelos de Riscos Proporcionais , RNA Viral/sangue , Ativação Viral
14.
Clin Immunol ; 229: 108773, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34102315

RESUMO

BACKGROUND: The dynamics of viral reservoir decay and naïve CD4 T-cell recovery between immunological non-responders (INR) and complete responders (CR) during long-term antiretroviral treatment (ART) are not fully known. METHODS: Twenty-eight chronic HIV-infected individuals on 5-year ART were divided into two groups: INR (CD4 counts ≤350 cells/µL, n = 13) and CR (CD4 counts ≥500 cells/µL, n = 15). The levels of HIV DNA and cell-associated HIV RNA (CA-RNA), CD4 counts, naïve CD4 counts and their correlations were analyzed at baseline, years 1, 3 and 5 of ART between the two groups. Expression of PD-1 on CD4 T-cells was quantified by flow cytometry. Linear mixed effect models were used to estimate the change procession in repeated measurements over 5 years. Slopes of the above-mentioned indicators were estimated using participant-specific linear regressions, respectively. RESULTS: INR maintained higher levels of HIV DNA and CA-RNA with higher percentages of PD-1+CD4 T-cells compared with CR during 5-year ART, concurrent with lower naïve CD4 T-cells. However, the rates of HIV DNA and CA-RNA decay in INR were not different from that in CR over time, and INR had higher rates of naïve CD4 T-cell percentage recovery. The baseline levels of HIV DNA were positively associated with the 5-year levels of HIV DNA, but negatively associated with the 5-year naïve CD4 counts. CONCLUSIONS: INR maintained significantly higher viral reservoir and lower naïve CD4 T-cells compared with CR during 5-year ART, however, the rates of reservoir decay and naïve CD4 T-cell percentage growth within INR were not lower than that in CR over time.


Assuntos
Terapia Antirretroviral de Alta Atividade , Linfócitos T CD4-Positivos/imunologia , Infecções por HIV/imunologia , Infecções por HIV/virologia , Adulto , Contagem de Linfócito CD4 , China , DNA Viral/sangue , DNA Viral/genética , Progressão da Doença , HIV/efeitos dos fármacos , HIV/genética , Infecções por HIV/tratamento farmacológico , Sobreviventes de Longo Prazo ao HIV , Humanos , Modelos Lineares , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , RNA Viral/sangue , RNA Viral/genética , Fatores de Tempo , Carga Viral/efeitos dos fármacos
15.
Arch Virol ; 166(9): 2435-2442, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34146173

RESUMO

Little is known about the usefulness of saliva samples for hepatitis B virus (HBV) genotyping and mutation analysis. The aim of this study was to evaluate the usefulness of oral fluid samples to determine HBV genotype distribution, S/polymerase mutations, and HBV subpopulation diversity among chronically HBV-infected individuals. Serum and oral fluid samples were obtained from 18 individuals for PCR and nucleotide sequencing of the HBV surface antigen gene. Biochemical analysis of liver enzymes (ALT, AST, GGT) and HBV, HCV, and HIV serological tests were also performed. All serum samples were HBsAg (+), anti-HBc (+), and anti-HBs (-); 55.6% were HBeAg (+)/anti-HBe (-), and 11.1% were anti-HIV (+). The mean HBV DNA viral load was 6.1 ± 2.3 log IU/mL. The HBV genotype distribution was as follows: A, 72.2%; D, 11.1%; E, 5.6%; F, 11.1%. A concordance of 100% in genotype classification and 99.8% in sequence similarity between paired oral fluid and serum samples was observed. HBsAg mutations were detected in all samples, but no resistance mutations were found in the polymerase gene. This study demonstrates that oral fluid samples can be used reliably for tracking HBV mutations, genotyping, and phylogenetic analysis. This could be important for molecular epidemiology studies with hard-to-reach populations.


Assuntos
Genótipo , Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Mutação , Filogenia , Adulto , Sequência de Bases , DNA Viral/sangue , DNA Viral/genética , Feminino , Hepatite B/virologia , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Antígenos E da Hepatite B/sangue , Hepatite B Crônica/virologia , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Testes Sorológicos
16.
BMC Cancer ; 21(1): 651, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-34074258

RESUMO

OBJECTIVE: Plasma Epstein-Barr virus (EBV) DNA is considered a biomarker for nasopharyngeal carcinoma (NPC). However, its long-term role in NPC development is unclear. MATERIALS AND METHODS: A total of 1363 participants seropositive for EBV VCA-IgA and EBNA1-IgA in a community-based NPC screening program in southern China were tested for plasma EBV DNA levels by real-time qPCR between 2008 and 2015. New NPC cases were confirmed by active follow-up approach and linkage to local cancer registry through the end of 2016. Cox proportional hazards regression analysis was performed to calculate the hazard ratios (HRs) for NPC risk with plasma EBV DNA. RESULTS: Thirty patients were newly diagnosed during a median 7.5 years follow-up. NPC incidence increased with the plasma EBV DNA load ranging from 281.46 to 10,074.47 per 100,000 person-years in participants with undetectable and ≥ 1000 copies/ml levels; the corresponding cumulative incidence rates were 1.73 and 50%. Furthermore, plasma EBV DNA loads conferred an independent risk for NPC development after adjustment for other risk factors, with HRs of 7.63 for > 3-999 copies/ml and 39.79 for ≥1000 copies/ml. However, the HRs decreased gradually after excluding NPC cases detected in the first 2 to 3 years and became statistically nonsignificant by excluding cases detected during the first 4 years. CONCLUSION: Elevated plasma EBV DNA can predict NPC risk over 3 years. Monitoring plasma EBV DNA can be used as a complementary approach to EBV serological antibody-based screening for NPC.


Assuntos
Biomarcadores Tumorais/sangue , DNA Viral/sangue , Infecções por Vírus Epstein-Barr/epidemiologia , Carcinoma Nasofaríngeo/epidemiologia , Neoplasias Nasofaríngeas/epidemiologia , Adulto , Idoso , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Infecções por Vírus Epstein-Barr/sangue , Infecções por Vírus Epstein-Barr/diagnóstico , Infecções por Vírus Epstein-Barr/virologia , Feminino , Herpesvirus Humano 4/genética , Herpesvirus Humano 4/imunologia , Herpesvirus Humano 4/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/sangue , Carcinoma Nasofaríngeo/diagnóstico , Carcinoma Nasofaríngeo/virologia , Neoplasias Nasofaríngeas/sangue , Neoplasias Nasofaríngeas/diagnóstico , Neoplasias Nasofaríngeas/virologia , Estudos Prospectivos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Risco , Testes Sorológicos/estatística & dados numéricos
17.
Gac Med Mex ; 157(1): 35-40, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34125813

RESUMO

INTRODUCTION: Identification of hepatitis B virus carriers in blood donors is imperative in order to avoid transmission of the disease via blood transfusion. OBJECTIVE: To determine if blood donors with positive results for serological markers HBsAg and anti-HBc were hepatitis B virus DNA carriers. METHODS: 12,745 samples were collected from six Ecuadorian blood banks and analyzed for HBsAg, anti-HBc and anti-HBs infectious markers by automated ELISA. All samples that tested positive for one, two or all three markers were analyzed with molecular techniques to determine the presence of viral DNA. RESULTS: 27.5 % of the samples that were reactive for anti-HBc alone and 100 % of those with positive results for HbsAg and IgM/IgG anti-HBc were identified to contain hepatitis B virus DNA (p = 0.001). CONCLUSIONS: The selection of infection markers, as well as the detection methods define the results. Performing two serological and one molecular test is important in order to identify hepatitis B virus carriers and prevent its transmission.


Assuntos
Doadores de Sangue , DNA Viral/sangue , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Biomarcadores/sangue , Bancos de Sangue , Doadores de Sangue/estatística & dados numéricos , Portador Sadio/diagnóstico , Portador Sadio/virologia , Equador , Ensaio de Imunoadsorção Enzimática/métodos , Vírus da Hepatite B/imunologia , Humanos
18.
Transfusion ; 61(8): 2240-2244, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34056747

RESUMO

BACKGROUND: Human parvovirus B19 (B19) is a pathogen that threatens the quality of plasma products. Therefore, health authorities have mandated measures against B19 contamination of plasma pools. The US FDA has recommended a B19 genome level of 104 IU/ml or lower in pooled plasma lots. Therefore, the B19 nucleic acid amplification test (B19-NAT) has been introduced in many plasma fractionators. However, in the Japanese Red Cross, which is the only approved blood collector in Japan, the B19 antigen test has been introduced for screening donated blood in Japan. Therefore, to clarify whether the antigen test is robust enough to screen blood samples according to the FDA recommendation, we evaluated B19 genome levels in each pooled plasma lot from 2003 to 2020. STUDY DESIGN AND METHODS: Data of 5576 pooled plasma lots from factories A and B, which were derived from plasma bags and passed the B19 antigen-based tests, receptor-mediated hemagglutination assay (B19-RHA), or chemiluminescent enzyme immunoassay (B19-CLEIA), during 2003 to 2020, were evaluated. The amount of B19 genome in each lot was determined using quantitative or semiquantitative B19-NAT. RESULTS: The B19 genome levels in pooled plasma lots screened using B19-RHA did not meet the FDA recommendation, whereas the lots derived from B19-CLEIA fulfilled the FDA recommendation, even during the B19 epidemic in Japan. DISCUSSION: The results suggest that the B19-CLEIA donor screening for plasma pools is also useful in light of the US FDA recommendation.


Assuntos
Antígenos Virais/análise , DNA Viral/análise , Infecções por Parvoviridae/diagnóstico , Parvovirus B19 Humano/isolamento & purificação , Antígenos Virais/sangue , DNA Viral/sangue , Humanos , Técnicas Imunoenzimáticas/métodos , Programas de Rastreamento , Técnicas de Amplificação de Ácido Nucleico/métodos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/virologia
19.
Eur J Cancer ; 151: 63-71, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33964573

RESUMO

BACKGROUND: To investigate the value of post-induction chemotherapy (IC) cell-free Epstein-Barr virus DNA (cfEBV DNApostIC) for prognostication in locally advanced nasopharyngeal carcinoma (LA-NPC). METHODS: A total of 910 histologically proven LA-NPC undergoing radical IC + concurrent chemo-radiotherapy (CCRT) or targeted radiotherapy (CTRT) or both (CTCRT) were involved. The concentration of cfEBV DNA was measured by quantitative polymerase chain reaction pre-IC (cfEBV DNApreIC) and at IC completion. CfEBV DNApostIC was classified as undetectable (0 copy/ml) and detectable (>0 copy/ml). Recursive partitioning analysis (RPA) with respect to the overall survival (OS) was applied to construct a risk stratification system incorporating cfEBV DNApostIC and critical risk factors. RESULTS: We observed that 660 (72.5%) and 250 (27.5%) patients had cfEBV DNApostIC undetectable and detectable respectively. CfEBV DNApostIC positive was associated with a significant inferior 5-year OS (76.2% versus 85.9%), metastasis-free survival (DMFS, 71.7% versus 86.4%) and disease-free survival (DFS, 57.7% versus 80.1%) than cfEBV DNApostIC negative (P < 0.001 for all). Additionally, cfEBV DNApostIC was independently significant for OS (hazard ratio [HR] 1.90, 95% CI 1.40-2.59), DMFS (1.99, 1.45-2.71) and DFS (2.38, 1.86-3.06) in multivariate analyses (P < 0.001 for all). RPA modelling yielded three distinct risk groups: low-risk (N0-1 and undetectable cfEBV DNApostIC or N2-3 and pre-treatment cfEBV DNA [cfEBV DNApreIC] <7000), median-risk (N0-1 and detectable cfEBV DNApostIC or N2-3 and cfEBV DNApreIC ≥7000 with undetectable cfEBV DNApostIC) and high-risk (N2-3 and cfEBV DNApreIC ≥7000 with detectable cfEBV DNApostIC), with 5-year OS of 88.1%, 79.2% and 66.9%, respectively. Our risk stratification outperformed TNM classification for predicting death (AUC, 0.631 versus 0.562; P = 0.012) and distant metastasis (0.659 versus 0.562; P = 0.004). CONCLUSIONS: CfEBV DNApostIC represents an effective indicator of prognostication in LA-NPC. We developed a risk classification system that provides improved OS prediction over the current staging system by combining cfEBV DNApostIC, cfEBV DNApreIC and N-stage classification in LA-NPC.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Ácidos Nucleicos Livres/sangue , DNA Viral/sangue , Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/genética , Quimioterapia de Indução , Carcinoma Nasofaríngeo/tratamento farmacológico , Neoplasias Nasofaríngeas/tratamento farmacológico , Adulto , Protocolos de Quimioterapia Combinada Antineoplásica/efeitos adversos , Quimiorradioterapia , Bases de Dados Factuais , Intervalo Livre de Doença , Infecções por Vírus Epstein-Barr/sangue , Infecções por Vírus Epstein-Barr/mortalidade , Feminino , Humanos , Quimioterapia de Indução/efeitos adversos , Quimioterapia de Indução/mortalidade , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/mortalidade , Carcinoma Nasofaríngeo/secundário , Carcinoma Nasofaríngeo/virologia , Neoplasias Nasofaríngeas/mortalidade , Neoplasias Nasofaríngeas/patologia , Neoplasias Nasofaríngeas/virologia , Estadiamento de Neoplasias , Intervalo Livre de Progressão , Radioterapia de Intensidade Modulada , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Fatores de Tempo , Carga Viral
20.
Sci Rep ; 11(1): 10292, 2021 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-33986444

RESUMO

Highly pathogenic bovine papillomaviruses (BPVs) were detected and quantified for the first time using digital droplet polymerase chain reaction (ddPCR) by liquid biopsy in 103 clinically healthy sheep. Overall, ddPCR detected BPVs in 68 blood samples (66%). BPV infection by a single genotype was revealed in 61.8% of the blood samples, and BPV coinfection by double, triple or quadruple genotypes was observed in 38.2% of liquid biopsies. The BPV-2 genotype was most frequently seen in sheep, whereas BPV-1 was the least common. Furthermore, ddPCR was very useful for detection and quantification; the BPV-14 genotype was observed for the first time in ovine species, displaying the highest prevalence in some geographical areas (Apulia). In 42 of the positive samples (61.8%), a single BPV infection was observed, 26 of which were caused by BPV-2 (61.9%) and 7 by BPV-13 (16.7%). BPV-14 was responsible for 7 single infections (16.7%) and BPV-1 for 2 single infections (4.7%). Multiple BPV coinfections were observed in the remaining 26 positive samples (38.2%), with dual BPV-2/BPV-13 infection being the most prevalent (84.6%). BPV infection by triple and quadruple genotypes was also observed in 11.5% and 3.8% of cases, respectively. The present study showed that ddPCR, a biotechnological refinement of conventional PCR, is by far the most sensitive and accurate assay for BPV detection compared to conventional qPCR. Therefore, ddPCR displayed an essential diagnostic and epidemiological value very useful for the identification of otherwise undetectable BPV genotypes as well as their geographical distributions and suggesting that animal husbandry practices contribute to cross-species transmission of BPVs.


Assuntos
Papillomavirus Bovino 1/genética , DNA Viral/sangue , Reação em Cadeia da Polimerase/veterinária , Ovinos/virologia , Animais , Bovinos , Genes Virais , Genótipo , Biópsia Líquida , Reação em Cadeia da Polimerase/métodos
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