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A small percentage of couples who regularly donated blood in China tested positive for HBsAg. Although it is well known that blood donors can acquire hepatitis B virus (HBV) infection from a chronically infected sexual partner, the prevalence of occult hepatitis B infections (OBIs) among blood donations from partners of HBV-infected chronically infected spouses and the risk to blood safety remain poorly understood. Among 212 763 blood donors, 54 pairs of couples (108 donations) were enrolled because one partner tested positive for HBsAg. Several molecular and serological examinations were conducted. The origin of HBV transmission between sexual partners was investigated further. Also evaluated was the potential risk of HBV infection with OBIs. We identified 10 (10/54, 18.6%) sexual partners of chronically infected HBV donors who were positive for HBV DNA, including five samples (9.3%) with OBIs, of which 3 (3/54, 5.6%, 1 in 70 921 donations) passed the routine blood screening tests. Seven of the 10 HBV-DNA-positive couples contracted the virus possibly through sexual or close contact. Among infected couples, immune escape mutations were observed. A high prevalence of OBIs was found among the partners of chronically infected HBV blood donors, posing a potential threat to blood safety.
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Doadores de Sangue , Segurança do Sangue , Hepatite B , Cônjuges , Segurança do Sangue/estatística & dados numéricos , Hepatite B/epidemiologia , Hepatite B/prevenção & controle , Hepatite B/transmissão , Cônjuges/estatística & dados numéricos , Prevalência , China/epidemiologia , Doadores de Sangue/estatística & dados numéricos , Vírus da Hepatite B , Humanos , Masculino , Feminino , Adulto Jovem , Adulto , Pessoa de Meia-IdadeRESUMO
BACKGROUND: In China, the vaccinated blood donors have rapidly increased by recent years, which may impact blood safety. The true prevalence of HBV between vaccinated blood donors and non-vaccinated blood donors should be explored. STUDY DESIGN AND METHODS: The samples of blood donors were collected and detected for serologic markers of HBV in the Shenzhen Blood Centre (SZBC). The discrepant results were tested with commercial electrochemiluminescence immunoassay (ELCI) for HBsAg, anti-HBs, HBeAg, Anti-HBe and Anti-HBc, alternative MPX ID NAT, nested PCR, and a quantitative real-time polymerase chain reaction (qPCR) assay for HBV DNA. The serological and molecular characteristics of HBV infected blood donors were analysed, and the effects on blood safety for donors born before and after the implementation of universal HBV vaccination were compared. RESULTS: Out of 242 presumed HBV infected donors from 26 318 donations, 131 (0.49%, [95% CI, 0.43-0.59]) chronic HBV infections (CHB, HBsAg detected with or without DNA), 58 (0.22%, [95% CI, 0.17-0.28]) occult hepatitis B infections (OBI, HBsAg not detected, assume anti-HBc positive and/or anti-HBs with HBV DNA) and 3 (0.011%, [95% CI, 0.0023-0.033]) window period (WP) infections were confirmed respectively. There were 28 CHBs (0.44%), 7 OBIs (0.11%) and 1 WP (0.016%) from vaccinated blood donor and 103 CHBs (0.52%), 51 OBIs (0.26%) and 2 WPs (0.01%) from non-vaccinated blood donor. The HBV+ (CHBs, OBIs and WPs) rate (0.56%) in vaccinated donors was lower than in non-vaccinated donors (0.78%, p < 0.05). The HBsAg titers of vaccinated infected blood donors (Median: 128.8 IU/ml) were much higher than non-vaccinated infected blood donors (58.4 IU/ml). The OBI yield rates in the vaccinated blood donors was significantly lower than the non-vaccinated blood donors (p < 0.05). There 102/124 (82.3%) samples were genotype B, 22/124 (17.7%) were genotype C respectively. There was no significant difference in the distribution of genotype between non-vaccinated blood donors (B/C, 86/17) and vaccinated blood donors (B/C, 23/6; p > 0.05). High frequency of vaccine escape mutations M133L (32.4%) and E164G in S region of genotype B strains and substitution L175S (40.9%) related to vaccine escape in S region of genotype C strains were identified. CONCLUSION: The universal HBV vaccination program markedly reduces the risk of HBV infection in blood donors, and provides a significant guarantee for the safety of blood transfusion. Several important mutations detected related vaccine escape and notable mutations needed further investigated.
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Doadores de Sangue , Vírus da Hepatite B , Hepatite B , Humanos , China/epidemiologia , DNA Viral , Hepatite B/epidemiologia , Anticorpos Anti-Hepatite B , Antígenos de Superfície da Hepatite B , Vacinas contra Hepatite B , Vírus da Hepatite B/genética , Reação em Cadeia da Polimerase em Tempo Real , VacinaçãoRESUMO
We report on a laser-diode (LD)-pumped master-oscillator power amplifier (MOPA) mid-infrared laser system based on an LD side-pumped Er:YSGG seed laser that can operate in both free-running and Q-switched regimes. In the free-running mode of the seed laser, the maximum amplified single-pulse energy was 83.4 mJ. In Q-switched mode of the seed laser, a maximum single-pulse energy of 7.8 mJ was achieved at 100 Hz repetition rate with the pulse width of 90 ns, corresponding to the peak power of 86.7 kW and the single-pass amplification factor of 1.66. The results indicate that the LD side-pumped MOPA structure is an effective way to realize a nanosecond â¼3µm mid-infrared laser with high repetition rate and high pulse energy.
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We report on a quasi-continuous Er:YAG planar waveguide laser operated at 2.94 µm based on the major oscillator power amplification configuration. With the total pump peak power of 32.01 kW, a maximum output peak power of 1.14 kW was obtained at the seed injection peak power of 184.4 W operated at 400µs, 40â Hz. Furthermore, the numerical simulation results indicate that better performance of the laser could be obtained with the higher injected seed laser power. To the best of our knowledge, this is the first experimental demonstration of 2.94 µm planar waveguide laser with an Er doped host material.
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BACKGROUND: Hepatitis B virus (HBV) infection is a major concern for blood safety in high-prevalence HBV countries such as China. In Shenzhen, dual hepatitis B surface antigen (HBsAg) enzyme-linked immunosorbent assays (ELISAs) have been adopted in parallel with nucleic acid testing (NAT) for donors for over a decade. A small proportion of blood donors test reactive (R) for HBsAg but negative through routine NAT, which can lead to HBV infection with an extremely low viral load. OBJECTIVES: We aimed to investigate and analyze the molecular characteristics of HBV among blood donors that tested HBsAg R in a single ELISA test. METHODS: Blood donations were evaluated in this study if confirmed HBsAg R through one of two ELISA kits. Samples with non-reactive (NR) results by NAT were collected and tested for HBsAg by chemiluminescent microparticle immunoassay (CLIA) with a neutralization test. The level of HBsAg was further assessed by electrochemiluminescence immunoassay (ECLIA). The viral basic core promoter (BCP) and pre-core (PC) and S regions were amplified by nested PCR. Quantitative real-time PCR (qPCR) for viral load determination and individual donation (ID)-NAT were adopted simultaneously. HBsAg was confirmed with CLIA, ECLIA, nested PCR, qPCR, and ID-NAT. RESULTS: Of the 100,252 donations, 38 and 41 were identified as HBsAg R with Wantai and DiaSorin ELISA kits, respectively. Seventy-nine (0.077%, 79/100,252) blood samples with ELISA R-NR and NAT NR results were enrolled in the study. Of these, 17 (21.5%,17/79) were confirmed as HBsAg-positive. Of the 14 genotyped cases, 78.6% (11/14) were genotype B, and C and D were observed in two and one sample, respectively. Mutations were found in the S gene, including Y100C, Y103I, G145R, and L175S, which can affect the detection of HBsAg. A high-frequency mutation, T1719G (93.3%), was detected in the BCP/PC region, which reduced the viral replication. CONCLUSION: A small number of blood samples with HBsAg ELISA R-NR and NAT NR results were confirmed as HBV infection, viral nucleic acids were found in most of the samples through routine NAT methods. It is necessary to employ more sensitive and specific assays for the detection of HBV infection among blood donors.
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Doadores de Sangue , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Hepatite B , China , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Técnicas de Amplificação de Ácido Nucleico , Filogenia , Reação em Cadeia da Polimerase/métodosRESUMO
BACKGROUND: Hepatitis B virus (HBV) infection is one of the major concerns for the safety of blood transfusion in high-prevalent countries such as in China. Prior studies outside of China have shown hepatitis B surface antigen (HBsAg) false-reactive rate of 0.02% to 0.04%. Similarly, false-negative HBsAg and HBV DNA results may occur in infected donors. Our study analyzed HBsAg enzyme-linked immunosorbent assay (ELISA)-reactive but NAT-negative donations in Shenzhen Blood Center, China. STUDY DESIGN AND METHODS: HBsAg ELISA-positive/NAT-negative plasma samples identified from screening 101,025 donations during 2017-2018 were analyzed by molecular and serologic tests including neutralization, chemiluminescence immunoassays, and various HBV DNA amplification assays. Molecular characterizations of HBsAg-positive/NAT-negative samples were determined by quantitative polymerase chain reaction (qPCR) and nested PCR amplification of the basic core and precore promotor regions (295 base pairs) and HBsAg (S) region (496 base pairs). RESULTS: Screening of 101,025 eligible blood donations identified 157 (0.16%, 95% confidence interval, 0.13%-0.18%) HBsAg ELISA-positive/NAT-negative plasma samples; of those, 71 (45.2%) were HBsAg confirmed positive by further HBsAg testing and DNA positive by molecular tests with increased sensitivity. Of the 71, all but one was antibody to hepatitis B core antigen reactive without antibody to hepatitis B surface antigen, yielding one recent (window-period) HBV infection. Of the remaining donations, 80 (51%) were not considered as HBV-infected donors, and 6 (3.8%) were interpreted as indeterminate since HBsAg results were discordant with unconfirmed HBV DNA results. In the 71 confirmed positives, HBsAg levels ranged from 0.05 to 400 IU/mL and HBV DNA from 6 to 2654 IU/mL; however, the correlation between the two was weak (R2 = 0.24). CONCLUSION: Fewer than half of HBsAg ELISA-positive/NAT-negative samples were confirmed as HBsAg positive. Our study demonstrates that in highly HBV-endemic countries, assays with high sensitivity and specificity may be required.
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Doadores de Sangue , DNA Viral/sangue , Seleção do Doador , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/metabolismo , Hepatite B/sangue , Adulto , China , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: Blood donor plasma samples were detected by the Ultrio Plus NAT system for HBV, HCV and HIV-1 in Shenzhen blood center, China. Reactive samples underwent further discriminatory testing of a single virus by the same methodology. A large number of cases of non-discriminated reactive (NDR) donors were found, leaving potential risk of transmitting HBV if not deferrals. This study identified those non-discriminated samples. METHODS: The NDR plasma samples from blood donation screening were detected and classified by additional molecular and serological tests. Molecular characterizations of DNA+ NDR were determined by sequencing analysis. RESULTS: A number of 259 (0.21%) NDR plasma samples from screening of 123,280 eligible blood donors were detected, which presented a higher rate (91.1%) of anti-HBc reactivity and nearly half (46.7%) of HBV DNA+ that classified as occult HBV infection (OBI). Most OBI strains were wild-type HBV, but some substitutions V168A, S174 N, V177A, Q129R/L/H, G145A/R in S region of genotype B (OBIB) and T47K/V/A, P49H/L, Q101R/H/K, S174 N, L175S, V177A, T118 M/R/K, G145R/A/K/E, R160K/N in S region of genotype C (OBIC) strains were identified in high frequency. CONCLUSION: Nearly half of NDR blood samples were identified as OBI, in which a number of important mutations were detected. NDR donation might have potential risk for HBV transmission, but need to be further investigated.
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Doadores de Sangue , Vírus da Hepatite B/genética , Hepatite B/virologia , Adulto , China , Feminino , Genótipo , HIV-1/genética , Hepacivirus/genética , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Humanos , Masculino , Mutação , Reação em Cadeia da Polimerase/métodos , Testes Sorológicos/métodosRESUMO
We designed a high-slope-efficiency and high-power laser diode (LD) side-pump Er:YSGG laser based on the analysis for the effect of the crystal dimension and the LD distribution on the temperature and energy distributions in laser crystal. A maximum output power of 34.9 W for a 2.8 µm mid-infrared laser was achieved at 200A, 120 Hz, and 500 µs pulse width, corresponding to the slope efficiency of 13.7% and optical-optical efficiency of 12.7%. Moreover, the beam quality $M_x^2/M_y^2$Mx2/My2 factors of the laser were measured to be 5.15/5.19 and the far-field divergences ${\Theta _x}/{\Theta _y}$Θx/Θy were 9.52/9.75 mrad.
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BACKGROUND: Neonatal hepatitis B vaccination program at birth has been implemented nationwide since 1992 in China. However, current HBV prevalence status in blood donors has not been entirely examined, which may impact HBV safety in blood donations as the vaccinees over 18 years old progressively become the majority population of blood donors. METHODS: In this study, 569,145 blood donors were screened for HBsAg by rapid tests and enzyme immunoassays, among them 475,538 blood samples with negative HBsAg were further screened for HBV DNA by nucleic acid testing between 2005 and 2014 at Shenzhen blood center. RESULTS: An overall 2.3 % HBsAg prevalence was found in the blood donor population during the past 10 years (2.86 % in 2005, 1.76 % in 2010, and 2.79 % in 2014, respectively). HBsAg seroconversion occurred in 0.37 % of repeat-donors. When stratified by age, the prevalence of HBsAg was found significantly higher in younger donors age 18-25 years (2.73 %) than in those 26-35 years (2.13 %), 36-45 years (2.03 %) and 46-58 years (1.71 %) (P < 0.001), unexpectedly suggesting that younger donors remained at risk of chronic HBV infection. Assuming that donors aged 18-22 born before or after 1992 were non-vaccinated and vaccinated, respectively, HBsAg prevalence was higher in first-time donors born ≥1992 (3.9 %) than prior to 1992 (3.5 %, P = 0.005). The incidence of HBV infection in the 5-year period examined was significantly lower in repeat-donors born ≥1992 (0.27 %) than prior to 1992 (0.6 %, P = 0.008). The yield of HBV DNA+/HBsAg- donors was 1:3,302, including 1:4,486 occult infections and 1:43,231 window period infections. CONCLUSION: Young blood donors born after implementation of universal HBV vaccination in China presented higher prevalence of HBsAg but lower incidence of HBsAg seroconversion than older, presumed unvaccinated, donors. HBV vaccine boosting for adolescents at 15-17 years old prior to reaching blood donor age might help improve blood safety.
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Doadores de Sangue , Antígenos de Superfície da Hepatite B/sangue , Vacinas contra Hepatite B/administração & dosagem , Adolescente , Adulto , Segurança do Sangue , China/epidemiologia , Doença Crônica , Feminino , Hepatite B/epidemiologia , Vírus da Hepatite B/imunologia , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico , Prevalência , Adulto JovemRESUMO
BACKGROUND: In East Asia, individuals systematically vaccinated at birth to hepatitis B virus (HBV) are an increasing part of the blood donor population. Their environment presents a high risk of contact with HBV. HBV vaccine efficacy and potential safety risk carried by vaccinated donors were examined. STUDY DESIGN AND METHODS: A total of 2028 vaccinated blood donors were recruited in 2012 and 2013 and tested for serologic (hepatitis B surface antigen [HBsAg], antibody to hepatitis B surface antigen [anti-HBs], and antibody to hepatitis B core antigen [anti-HBc]) and molecular (HBV DNA) markers of HBV. HBsAg, anti-HBs, and viral load were quantified. RESULTS: Donors 18 to 21 years systematically vaccinated at birth and 22 to 25 years and older donors had both 30.0% negative serology and 1.8% anti-HBc only but the latter group carried significantly higher prevalence of anti-HBc (p < 0.0001). Anti-HBc, mostly associated with anti-HBs, increased from 10.7% at age 18 to 31.5% at age 25. The level of anti-HBs was significantly higher in anti-HBc-positive donors than in anti-HBs-only donors (p < 0.0001). Samples from 24 donors contained low viral load (25 ± 22 IU/mL), half of them undetected by standard nucleic acid testing (NAT), and were classified as four recent infections, 17 occult HBV infections (OBI), and three primary OBIs. Eighteen of 24 carried anti-HBs; 14 of 15 strains were wild-type Genotype B and one was Genotype C. CONCLUSIONS: In an environment of frequent high Genotype B or C viremia, blood donors vaccinated at birth are frequently but mildly infected: asymptomatic and normal alanine aminotransferase level, identified by anti-HBc seroconversion and boosting of anti-HBs. Low viral load and frequent anti-HBs limit transfusion risk.
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Doadores de Sangue , Segurança do Sangue , Anticorpos Anti-Hepatite B/sangue , Antígenos do Núcleo do Vírus da Hepatite B/imunologia , Vacinas contra Hepatite B , Vírus da Hepatite B/isolamento & purificação , Hepatite B/sangue , Vacinação , Viremia/sangue , Adolescente , Adulto , China/epidemiologia , DNA Viral/sangue , Suscetibilidade a Doenças , Feminino , Seguimentos , Hepatite B/epidemiologia , Hepatite B/imunologia , Hepatite B/prevenção & controle , Hepatite B/transmissão , Vírus da Hepatite B/genética , Vírus da Hepatite B/imunologia , Humanos , Masculino , Técnicas de Amplificação de Ácido Nucleico , Risco , Reação Transfusional , Carga Viral , Viremia/diagnóstico , Viremia/imunologia , Viremia/prevenção & controle , Viremia/transmissão , Adulto JovemRESUMO
A six-year pilot study on nucleic acid testing for HBV, HCV and HIV-1 has been undertaken on sero-negative plasmas in mini-pool and individual donation testing at Shenzhen Blood Center. Of 307,740 sero-negative blood samples, 95 of 102 HBV DNA yields were confirmed positive, 80/95 (84.2%) were classified as occult HBV infection (OBI) and 15 (15.8%) as window period cases. Amongst OBIs, 45% carried anti-HBc only, 41.3% anti-HBc and anti-HBs and 13.7% anti-HBs only. HBV DNA yield was 1:3239. One HCV WP and one HIV-1 infected donations were detected. High residual risk was found in current blood donations screening in China.
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Doadores de Sangue , DNA Viral/sangue , Seleção do Doador/métodos , HIV-1 , Hepacivirus , Vírus da Hepatite B , RNA Viral/sangue , China , Feminino , Humanos , Masculino , Projetos Piloto , Estudos RetrospectivosRESUMO
HLA-C*08:99 differs by one non-synonymous nucleotide from C*08:01:01 in exon 5, codon 288 GTT>ATT.
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Povo Asiático , Antígenos HLA-C , Alelos , Povo Asiático/genética , China , Éxons/genética , Antígenos HLA-C/genética , Sequenciamento de Nucleotídeos em Larga Escala , HumanosRESUMO
Immune control of various infectious diseases, particularly viral, was shown to be more efficient for females than males. Response to viral vaccines (HAV, HBV) was higher in females. Data on hepatitis B virus (HBV) markers accumulated over 15 years in blood donors was stratified according to sex, including HBsAg, HBV viral load and levels of anti-HBs in areas where genotypes B and C (China), genotype D (Iran, Lebanon, Tunisia) and genotype E (Ghana, Burkina Faso, Gabon) were prevalent. HBsAg was screened by either ELISA or rapid tests, anti-HBc and anti-HBs by ELISA, HBV DNA load by a standardized method across sites. In Ghanaian children less than 5 years, HBV DNA load was significantly lower in females than in males (p = 0.035). In China, Ghana, Burkina Faso and Gabon blood donors, median HBsAg prevalence was ~5% and 3% in China, ~8.5% and 4.5% in Gabon, ~16% and 11% in Burkina Faso and ~11% and 7% in Ghana for male and female donors, respectively (p < 0.001). In HBsAg+ Ghanaian blood donors, distribution and median viral load were not significantly different between sexes; occult hepatitis B infections (OBI) were significantly more frequent in males. In Chinese blood donor anti-HBc+ and anti-HBs+, anti-HBs levels tended to be higher in males but vaccinated donors' anti-HBs+ only, while anti-HBs levels were females > males. In areas where genotypes B-E are dominant, the prevalence of chronic HBV infection (HBsAg+) seems better controlled before age 16−18 by females infected vertically or horizontally. OBIs appear considerably more frequent in men, suggesting lower efficacy of HBV infection control. Female blood donors appear significantly safer from HBV than males, and their donation should be encouraged.
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Hepatite B Crônica , Hepatite B , Adolescente , Doadores de Sangue , Criança , DNA Viral/genética , Feminino , Gana/epidemiologia , Anticorpos Anti-Hepatite B , Antígenos do Núcleo do Vírus da Hepatite B , Antígenos de Superfície da Hepatite B , Vírus da Hepatite B/genética , Hepatite B Crônica/epidemiologia , Humanos , Masculino , Infecção Persistente , PrevalênciaRESUMO
Indoor environmental control is usually applied in poultry farming to ensure optimum growth conditions for birds. However, these control methods represent a considerable share of total energy consumption, and the trend of applying new equipment in the future for precision livestock farming would further increase energy demand, resulting in an increase in greenhouse gas emissions and management costs. Therefore, to ensure optimum efficiency of both energy use and livestock productivity, a customized hourly model was developed in the present study to interpret and analyze the electronically collected data. The modules for estimating indoor gas concentrations were incorporated into the present model, as this has not been properly considered in previous studies. A validation test was performed in a manure-belt layer house using sensors and meters to measure the indoor environmental parameters and energy consumption. The predicted results, including indoor temperature, relative humidity, carbon dioxide and ammonia concentrations, showed good agreement with the measured data, indicating a similar overall trend with acceptable discrepancies. Moreover, the corresponding differences between the measured and simulated energy consumption for heating, tunnel ventilation and base ventilation were 13.7, 7.5, and 0.1%, respectively. The total energy demand estimated by the model showed a limited discrepancy of approximately 10.6% compared with that measured in reality. Although human factors, including inspection, cleaning, vaccination, etc., were not included in the model, the validation results still suggested that the customized model was able to accurately predict the indoor environment and overall energy consumption during poultry farming. The validated model provides a tool for poultry producers to optimize production planning and management strategies, increase the production rate of unit energy consumption and achieve precision livestock farming from an energy consumption standpoint.
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Background: All Chinese blood centers have implemented mini pool (MP) HBV nucleic acid testing (NAT) together with HBsAg ELISA in routine donor screening since 2015. The prevalence of occult hepatitis B virus infection (OBI) in donors from different regions varies, and the molecular characterization of the HBV DNA and clinical outcomes of these OBIs remain largely unexplored. Methods: Blood donations from Heyuan city in Southern China were screened by HBsAg ELISA and HBV MP8 NAT. Donations with HBsAg-/HBV DNA+ were collected for this study. Molecular characterizations of HBV DNAs were further analyzed by various DNA amplification assays including quantitative PCR (qPCR) and nested PCR, amplifying the basic core and pre-core promoter regions (BCP/PC). The HBsAg (S) region from HBV DNA was isolated by high-volume nucleic acid extraction. Notable mutations were identified by comparison to the HBV reference sequences. The clinical outcomes of the donors with OBIs were further followed for nearly 3 years. Results: Seventy OBIs from 44,592 donations (0.15%) that we identified and reported previously were enrolled for this current study. HBV sequences were obtained from 44/70 OBIs, and genotyping analysis showed that 42/44 (95.2%) OBIs were genotype B, and 2/44 (4.8%) were genotype C. Interestingly, mutation analysis revealed that various mutations including M133L/T, F134L, P142L, V168A, R169H, S174N, L175S, and V177A of HBV DNA affecting HBsAg detection were observed in genotype B OBIs. Two notable mutations, T47K and L53S, were identified in genotype C OBIs. Follow-up studies showed that 3/31 (9.7%) OBIs converted to HBsAg+ as chronic infections while 1/31 (3.2%) HBV DNA was undetectable (classified as recovery) and 27/31 (87.1%) remained as OBIs. Conclusion: Various notable mutations affecting HBsAg detection were observed in blood donors with OBIs in Heyuan city of Southern China. Follow-up studies showed that most OBIs remained as OBIs with fluctuating or low viral loads. Higher sensitive HBV ID NAT is recommended for donor screening to further reduce the transmission risk of OBIs.
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Hepatite B Crônica , Hepatite B , Doadores de Sangue , DNA Viral/genética , Hepatite B/diagnóstico , Hepatite B/epidemiologia , Hepatite B/prevenção & controle , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/genética , Humanos , MutaçãoRESUMO
Prevalence and characteristics of occult hepatitis B virus (HBV) infection (OBI) of genotypes B and C prevalent in China have not been extensively explored. Characterization of OBI strains obtained from Chinese blood donors was based on clinical and serological analyses, follow-up testing, and sequence analyses. Twenty-eight samples from 165,371 HBV surface antigen (HBsAg)-negative plasmas were confirmed HBsAg negative and DNA positive(HBsAg(-)/DNA(+)), of which 22 were classified as OBIs and 6 as window period infections. The OBI incidence was 1:7,517 in blood donors, whose ages ranged between 20 and 45 years (median, 28 years). OBI donors had normal alanine aminotransferase (ALT) levels and low viral loads ranging between unquantifiable amounts and 178 IU/ml (median, 14 IU/ml). Sequences from 21 basic core promoter/precore (BCP/PC) regions, five whole genomes, and two additional pre-S/S regions from OBI strains were compared to genotypes B and C HBsAg(+) reference strains. Eighty-six percent (6/7) of OBI strains were genotype C. Deletions, insertions, stop codons, and substitutions were detected in 15/21 (71%) core regulatory elements of OBI strains. Critical mutations were found in the core proteins of 5/5 OBI strains in parallel with random substitutions in pre-S/S proteins from 6/7 (86%) OBI strains. Critical mutations in core regulatory elements and core proteins might affect OBI genotype B and C strain replication. That there were few S protein substitutions suggests a minor role of the host immune defenses in OBI occurrence.
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Doadores de Sangue , DNA Viral/sangue , Vírus da Hepatite B/isolamento & purificação , Hepatite B/diagnóstico , Hepatite B/epidemiologia , Adulto , China/epidemiologia , Análise por Conglomerados , DNA Viral/genética , Feminino , Genótipo , Hepatite B/patologia , Hepatite B/virologia , Anticorpos Anti-Hepatite B/sangue , Antígenos de Superfície da Hepatite B/sangue , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Prevalência , Análise de Sequência de DNARESUMO
Background: Most Chinese Blood Centers adopted mini pool (MP) nucleic acid testing (NAT) for HBV screening due to high cost of Individual donation (ID) NAT, and different proportions of MP-reactive but ID-non-reactive donations (MP+/ID-, defined as non-resolved donations) have been observed during daily donor screening process. Some of these non-resolved donations are occult HBV infections (OBIs), which pose potential risk of HBV transmission if they are not deferred. This study is aimed to further analyze these non-resolved donations. Methods: The non-resolved plasma samples were further analyzed by serological tests and various HBV DNA amplification assays including quantitative PCR (qPCR) and nested PCR amplifying the basic core and pre-core promoter regions (BCP/PC; 295 base pairs) and HBsAg (S) region (496 base pairs). Molecular characterizations of HBV DNA+ non-resolved samples were determined by sequencing analysis. Results: Of 17,226 MPs from 103,356 seronegative blood donations, 98 MPs were detected reactive for HBV. Fifty-six out of these 98 (57.1%) reactive MPs were resolved as HBV DNA+, but the remaining 42 pools (42.9%, 252 donations) were left non-resolved with a high rate (53.2%) of anti-HBc+. Surprisingly, among 42 non-resolved MPs, 17 contained one donation identified as OBIs by alternative NAT assays. Sequence analysis on HBV DNAs extracted from these OBI donations showed some key mutations in the S region that may lead to failure in HBsAg detection and vaccine escape. Conclusion: A total of 53.2% of the non-resolved donations were anti-HBc+, and OBIs were identified in 40.5% of these non-resolved pools. Therefore, non-resolved donations with anti-HBc+ might pose potential risk for HBV transmission. Our present analysis indicates that anti-HBc testing in non-resolved donations should be used to identify OBIs in order to further increase blood safety in China.
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Doadores de Sangue , DNA Viral/análise , Seleção do Doador/métodos , Antígenos do Núcleo do Vírus da Hepatite B/análise , Hepatite B , Adolescente , Adulto , China , Feminino , Hepatite B/diagnóstico , Hepatite B/epidemiologia , Vírus da Hepatite B , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico/métodos , Adulto JovemRESUMO
Rice-crayfish (Procambarus clarkii) coculture is an effective farming mode and has been promoted in various regions of China. However, infection in crayfish can be a significant economic drain. We found crayfish infected with Vibrio parahemolyticus (VP), and to understand the molecular mechanisms of the immune responses of crayfish to VP infection, Illumina sequencing was employed to identify changes in the mRNA of hepatopancreatic tissue. A total of 47.30 and 43.01million high-quality transcriptome reads were generated from the hepatopancreatic samples of the experimental group (EG) and control group (CG), respectively. We found 5559 genes were significantly differentially expressed, including 2521 up-regulated genes (45.35%) and 3038 down-regulated genes (54.65%). These genes were enriched in 126 GO terms and 76 KEGG pathways (Pâ¯≤â¯0.05), including the MAPK and PI3K-Akt signaling pathways and cell adhesion molecules, with 23 up-regulated genes and 3 down-regulated genes related to immune responses in the EG relative to the CG. Histopathological analysis revealed that the epithelial cells of the hepatopancreatic tubules in the EG were severely atrophic, necrotic, and exfoliated, resulting in thin and collapsing hepatopancreatic tubules. The expression patterns of 8 differentially expressed genes involved in immune responses were validated by quantitative real-time RT-PCR. These results provide a valuable basis for the immune responses of crayfish to acute hepatopancreatic necrosis disease at transcriptome level.
Assuntos
Proteínas de Artrópodes/genética , Astacoidea/genética , Astacoidea/imunologia , Hepatopâncreas/imunologia , Imunidade Inata/imunologia , Transcriptoma , Vibrio parahaemolyticus/fisiologia , Animais , Proteínas de Artrópodes/metabolismo , Astacoidea/microbiologia , Técnicas de Cocultura , Regulação da Expressão Gênica , Hepatopâncreas/metabolismo , Hepatopâncreas/microbiologia , Oryza/microbiologiaRESUMO
OBJECTIVES: Occult hepatitis B virus infection (OBI) carries a risk of hepatitis B virus (HBV) transmission and hepatocellular carcinoma. As previous studies have had a limited sample size, the characteristics of OBI with genotype B and C (OBIB and OBIC) mutations relating to hepatitis B surface antibody (anti-HBs) elicited by vaccination or a limited host immune response to HBV have not been fully explored. METHODS: In this study, the occurrence of OBIB or OBIC strains associated with envelope protein (pre-S/S) amino acid substitutions obtained from 99 blood donors stratified according to anti-HBs carriage were characterized extensively. RESULTS: According to the presence of anti-HBs within each genotype, the number and frequency of substitution sites specific for anti-HBs(-) OBIB were higher than those specific for anti-HBs(+) OBIB strains (67 vs 31; 117 vs 41), but the reverse pattern was found in OBIC strains (3 vs 24; 3 vs 26). Mutations pre-s1T68I and sQ129R/L were found uniquely in 15-25% of anti-HBs(+) OBIB carriers and mutation pre-s1A54E was found preferentially in anti-HBs(+) OBIC, while 17 substitutions were found preferentially in 11-38% of anti-HBs(-) OBIB strains. In the major hydrophilic region (MHR) region, mutations sS167 in OBIB, sT118 in OBIC, and sA166 in both genotypes were possibly immune-induced escape mutation sites. CONCLUSIONS: Several mutations in pre-S/S of OBI appeared to be associated with carrier anti-HBs pressure, which might be risk factors for potential reactivation of viruses under anti-HBs selection in OBI carriers.
Assuntos
Doadores de Sangue , DNA Viral/sangue , Antígenos de Superfície da Hepatite B/sangue , Vírus da Hepatite B/genética , Hepatite B/virologia , Mutação , Proteínas do Envelope Viral/genética , Adulto , Substituição de Aminoácidos , Carcinoma Hepatocelular/virologia , Feminino , Genótipo , Hepatite B/diagnóstico , Hepatite B/imunologia , Anticorpos Anti-Hepatite B/imunologia , Vírus da Hepatite B/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas do Envelope Viral/imunologia , Adulto JovemRESUMO
Leiocassis longirostris is a common fish variety that is widely cultivated in China, during the breeding process however, it is highly susceptible to bacterial haemorrhagic septicemia, which can cause great economic loss for farmers. To understand the immune responses of L. longirostris to Aeromonas hydrophila infection, Illumina sequencing was employed to identify changes in the mRNA and miRNA in spleen tissue. In this study, a total of 92.16 and 95.61 million (M) high-quality transcriptome reads were generated from the control group (CG) and experimental group (EG) spleen samples, respectively, and 207 up-regulated and 185 down-regulated genes were identified. These genes were enriched in 29 GO terms and 30 KEGG pathways (P ≤ 0.05), including cytokine-cytokine receptor interaction and complement and coagulation cascades, with 17 up-regulated genes and 12 down-regulated genes related to immune responses in the EG relative to the CG. Based on the zebrafish genome, miRNA-seq identified a total of 343 miRNAs, of which 15 were up-regulated and 10 were down-regulated (fold-change ≥2 or ≤0.5 and P ≤ 0.05). Target gene prediction and KEGG enrichment analysis revealed that all of the target genes were concentrated in 13 pathways associated with immune response, including the mTOR signaling pathway and the TGF-beta signaling pathway. The expression patterns of 8 differentially expressed genes and 4 miRNAs involved in immune response were validated by quantitative real-time RT-PCR. These results have provided valuable insights into the molecular mechanisms underlying the immune response of L. longirostris to bacterial haemorrhagic septicemia.