Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 242.809
Filtrar
1.
Z Gastroenterol ; 58(9): 868-871, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32947632

RESUMO

BACKGROUND: Concurrent cytomegalovirus (CMV) in inflammatory bowel disease (IBD)-related colitis is an important scenario associated with high rates of colectomy and other morbidity. Due to the low incidence of CMV, testing of all patients is associated with an unacceptably high consumption of resources and delay in treatment. Therefore, several predictive scores have been developed to identify patients at risk for a CMV infection. METHODS: We performed a retrospective single center study in a German University hospital including all IBD patients with available data on CMV-PCR analysis in whole blood between 2010 and 2018 and evaluated 2 prognostic scores for CMV infection for their diagnostic accuracy. RESULTS: In the study, 907 patients with IBD and CMV-PCR were identified. Of them, 21 patients (2.3 %) had a positive CMV-PCR (≥ 1000 copies/mL), 14 of them in ulcerative colitis and 7 in Crohn's disease. The Berlin Score identified 667 patients (73.1 %) as potentially CMV-positive, resulting in a positive predictive value of 2.5 % and a negative predictive value of 98.3 %. In contrast, the Münster Score identified 60 patients as potentially CMV-positive, resulting in a PPV of 20 % and an NPV of 99.4 %. CONCLUSIONS: Scoring systems can help to identify patients at risk for a CMV infection and minimize resource consumption and delay in treatment. Due to low incidence, a 2-step-algorithm, consisting of the Münster Score followed by a CMV-PCR if the score indicates a CMV infection, is preferable.


Assuntos
Infecções por Citomegalovirus/complicações , Infecções por Citomegalovirus/diagnóstico , Citomegalovirus/isolamento & purificação , Doenças Inflamatórias Intestinais/complicações , Infecções Oportunistas/complicações , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Colite Ulcerativa , Citomegalovirus/genética , Infecções por Citomegalovirus/sangue , DNA Viral/análise , Feminino , Humanos , Doenças Inflamatórias Intestinais/patologia , Masculino , Pessoa de Meia-Idade , Infecções Oportunistas/sangue , Infecções Oportunistas/patologia , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Estudos Retrospectivos , Sensibilidade e Especificidade
2.
Talanta ; 220: 121392, 2020 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-32928412

RESUMO

COVID-19 pandemic is a serious global health issue today due to the rapid human to human transmission of SARS-CoV-2, a new type of coronavirus that causes fatal pneumonia. SARS -CoV-2 has a faster rate of transmission than other coronaviruses such as SARS and MERS and until now there are no approved specific drugs or vaccines for treatment. Thus, early diagnosis is crucial to prevent the extensive spread of the disease. The reverse transcription-polymerase chain reaction (RT-PCR) is the most routinely used method until now to detect SARS-CoV-2 infections. However, several other faster and accurate assays are being developed for the diagnosis of COVID-19 aiming to control the spread of infection through the identification of patients and immediate isolation. In this review, we will discuss the various detection methods of the SARS-CoV-2 virus including the recent developments in immunological assays, amplification techniques as well as biosensors.


Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Técnicas Biossensoriais , Diagnóstico Precoce , Humanos , Imunoensaio , Pandemias , Reação em Cadeia da Polimerase
3.
Emerg Microbes Infect ; 9(1): 1955-1957, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32869728

RESUMO

The case refers to a 51-year-old symptomatic man with a new SARS-CoV-2 RNA positive nasopharyngeal swab after two negative ones and the lack of significant development of antibody response measured by different diagnostic serological test. Our case underlines that a discrepancy between clinical course of SARS-CoV-2 infection and results from diagnostic tests may exist. This concept is rapidly emerging and supports the need for a deep knowledge of available and "in development" tests for a correct interpretation of their findings.


Assuntos
Anticorpos Antivirais/sangue , Betacoronavirus/imunologia , Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/diagnóstico , Imunoglobulina G/sangue , Pneumonia Viral/diagnóstico , Infecções por Coronavirus/tratamento farmacológico , Ensaio de Imunoadsorção Enzimática , Reações Falso-Negativas , Humanos , Hidroxicloroquina/uso terapêutico , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/tratamento farmacológico , Reação em Cadeia da Polimerase , Quarentena , Recidiva , Testes Sorológicos , Carga Viral
4.
Rev Med Liege ; 75(9): 619-625, 2020 Sep.
Artigo em Francês | MEDLINE | ID: mdl-32909415

RESUMO

Tuberculosis (TB) is one of the top 10 causes of death worldwide. Abdominal TB is an uncommon presentation of TB and is challenging to diagnose due to its insidious onset. The most common forms in children are peritonitis and lymphadenitis. Diagnosis is based on radiological and histopathological findings. Specific PCR amplification confirms the diagnosis quicker than conventional cultures. The treatment includes a 6-month therapy and a close follow-up. This article describes the different methods allowing to confirm the diagnosis of abdominal TB.


Assuntos
Peritonite , Tuberculose , Criança , Humanos , Reação em Cadeia da Polimerase , Guias de Prática Clínica como Assunto , Radiografia
5.
Pneumologie ; 74(9): 615-620, 2020 Sep.
Artigo em Alemão | MEDLINE | ID: mdl-32916744

RESUMO

PATIENT HISTORY AND CLINICAL FINDINGS: A 46-year old construction worker presented at the emergency department with two orthostatic syncopes. The patient complained of prolonged fever and coughs for 7 days which had not improved after oral treatment with sultamicillin for 5 days, prescribed by the patient's general practitioner. Physical examination showed high blood pressure due to previously known hypertension. Other vital signs without pathological findings. Pulmonary auscultation showed basal soft crackling noises of the left lung. FINDINGS AND DIAGNOSIS: Laboratory examination showed increased values for LDH, pro-BNP and CRP and normal values for leucocytes and procalcitonin. Conventional X-Ray of the chest showed bipulmonal lateral atypical infiltrates. After the first PCR turned in negative another PCR-analysis for SARS-CoV-2 of a deep oral swab-sample was performed since the clinical, laboratory and radiological findings were typical for COVID-19. Again, SARS-CoV-2-RNA was not detected. A CT-scan of the chest showed bipulmonal lateral ground-glass attenuation, again typical for COVID-19 associated pneumonia. After a third attempt for a PCR-analysis of a deep oral swab-sample was negative, analysis of a sputum was performed which finally confirmed the diagnosis of COVID-19 associated pneumonia. THERAPY AND COURSE OF EVENTS: The patient was admitted for evaluation of syncopes and suspect of COVID-19 associated pneumonia. The patient was prophylactically isolated while the result of SARS-CoV-2-PCR from a deep oral swab was pending. Suspecting a possible secondary bacterial infection at the beginning, intravenous antibiotic treatment with ampicillin/sulbactam was initiated. While further examinations showed no indication for bacterial infection, antibiotics were discontinued after 3 days. Due to clinical recovery antiviral therapy was not performed after confirming the diagnosis. The patient was discharged 17 days after onset of first symptoms without any requirements for further isolation. CONCLUSION: This casuistic describes a case of COVID-19 associated pneumonia presenting with typical clinical features, laboratory and radiological findings. Detection of viral RNA was not successful from deep oral swab-samples despite repeated attempts. Finally, PCR-analysis of sputum confirmed the diagnosis. Analysis of deeper airway samples (sputum, bronchoalveolar lavage, tracheal secretions) or stool for SARS-CoV-2 should be performed in cases of evident clinical suspicion of COVID-19 and negative PCR results from deep oral swabs.


Assuntos
Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Betacoronavirus/isolamento & purificação , Técnicas de Laboratório Clínico , Humanos , Pulmão/diagnóstico por imagem , Pessoa de Meia-Idade , Orofaringe/virologia , Pandemias , Reação em Cadeia da Polimerase , RNA Viral/isolamento & purificação , Radiografia Torácica
6.
Acta Odontol Latinoam ; 33(2): 104-111, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32920612

RESUMO

Candida dubliniensis (Cd) and Candida albicans (Ca) are the most frequently isolated yeasts in HIV+ patients. Some of the enzymes produced by these yeasts are considered virulence factors since they contribute to pathogenicity of Candida spp. The aim of the present study was to compare production of enzymes such as phospholipase (Ph), proteinase (P), and hemolysin (H) by Cd and Ca strains isolated from periodontal HIV-positive patients receiving and not receiving highly active antiretroviral therapy (HAART). Subgingival biofilm samples were obtained using paper points, and a sample of oral mucosa was taken using a swab. Phenotypic and molecular methods were used to isolate 39 strains of Candida, including 25 strains of Cd and 14 strains of Ca, obtained from 33 periodontal pocket samples and 6 oral mucosa samples collected from 15 HIV+ patients (8 receiving and 7 not receiving HAART). Malt egg-yolk agar, albumin agar and blood agar were used to evaluate pH, P and H production respectively. The strains were inoculated in duplicate and incubated at 37 ºC. Colony and halo diameters were measured. A greater proportion of Ca was observed in patients not receiving HAART, and a higher proportion of Cd was observed in those under HAART, Chi2 p< 0.001. Phospholipase production was observed in 92.9% percent of isolated Ca strains but in none of the isolated Cd strains. Proteinase production was high in Ca and Cd strains isolated from patients not receiving HAART. Hemolysin production was observed in all the studied strains, though it was significantly higher (p=0.04) in Ca and Cd strains isolated from patients not receiving HAART. To sum up, the proportion of Candida dubliniensis strains was highest in the subgingival biofilm of patients receiving HAART, and Cd strains were found to express fewer virulence factors than Ca strains.


Assuntos
Terapia Antirretroviral de Alta Atividade/métodos , Biofilmes/crescimento & desenvolvimento , Candida albicans/enzimologia , Candida albicans/isolamento & purificação , Candida/enzimologia , Candida/isolamento & purificação , Candidíase Bucal/microbiologia , Gengiva/microbiologia , Infecções por HIV/complicações , Candida/classificação , Candida/genética , Candida albicans/genética , Candidíase Bucal/complicações , Genótipo , Infecções por HIV/microbiologia , Humanos , Mucosa Bucal/microbiologia , Fenótipo , Reação em Cadeia da Polimerase , Fatores de Virulência/genética
7.
Korean J Parasitol ; 58(4): 445-450, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32871638

RESUMO

Trombiculid "chigger" mites (Acari) are ectoparasites that feed blood on rodents and another animals. A crosssectional survey was conducted in 7 ecosystems of southern Vietnam from 2015 to 2016. Chigger mites were identified with morphological characteristics and assayed by polymerase chain reaction for detection of rickettsiaceae. Overall chigger infestation among rodents was 23.38%. The chigger index among infested rodents was 19.37 and a mean abundance of 4.61. A total of 2,770 chigger mites were identified belonging to 6 species, 3 genera, and 1 family, and pooled into 141 pools (10-20 chiggers per pool). Two pools (1.4%) of the chiggers were positive for Orientia tsutsugamushi. Ricketsia spp. was not detected in any pools of chiggers. Further studies are needed including a larger number and diverse hosts, and environmental factors to assess scrub typhus.


Assuntos
Orientia tsutsugamushi/isolamento & purificação , Roedores/parasitologia , Trombiculidae/microbiologia , Animais , Estudos Transversais , DNA Bacteriano/genética , Orientia tsutsugamushi/genética , Reação em Cadeia da Polimerase/métodos , Tifo por Ácaros/microbiologia , Trombiculidae/genética , Vietnã
8.
BMC Infect Dis ; 20(1): 648, 2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32883215

RESUMO

BACKGROUND: Due to the frequent reassortment and zoonotic potential of influenza A viruses, rapid gain of sequence information is crucial. Alongside established next-generation sequencing protocols, the MinION sequencing device (Oxford Nanopore Technologies) has become a serious competitor for routine whole-genome sequencing. Here, we established a novel, rapid and high-throughput MinION multiplexing workflow based on a universal RT-PCR. METHODS: Twelve representative influenza A virus samples of multiple subtypes were universally amplified in a one-step RT-PCR and subsequently sequenced on the MinION instrument in conjunction with a barcoding library preparation kit from the rapid family and the MinIT performing live base-calling. The identical PCR products were sequenced on an IonTorrent platform and, after final consensus assembly, all data was compared for validation. To prove the practicability of the MinION-MinIT method in human and veterinary diagnostics, we sequenced recent and historical influenza strains for further benchmarking. RESULTS: The MinION-MinIT combination generated over two million reads for twelve samples in a six-hour sequencing run, from which a total of 72% classified as quality screened, trimmed and mapped influenza reads to produce full genome sequences. Identities between the datasets of > 99.9% were achieved, with 100% coverage of all segments alongside a sufficient confidence and 4492fold mean depth. From RNA extraction to finished sequences, only 14 h were required. CONCLUSIONS: Overall, we developed and validated a novel and rapid multiplex workflow for influenza A virus sequencing. This protocol suits both clinical and academic settings, aiding in real time diagnostics and passive surveillance.


Assuntos
Sequenciamento de Nucleotídeos em Larga Escala/métodos , Vírus da Influenza A/genética , Sequenciamento por Nanoporos/métodos , Humanos , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Fluxo de Trabalho
9.
Medicine (Baltimore) ; 99(33): e21520, 2020 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-32871999

RESUMO

RATIONALE: Information regarding the clinical features and outcomes of pneumonia due to an infection with human coronavirus (HCoV)-OC43 in children with cancer is rare. This report presents the clinical features in terms of chest CT scan images which may be used to identify cases of HCoV-OC43 infection induced pneumonia in immunocompromised children. PATIENT CONCERNS: We report here a girl with acute lymphoblastic leukemia who developed respiratory symptoms during febrile neutropenia. Rapid clinical progression and nodular lesions on her chest X-ray and computed tomography scans were suggestive of a pulmonary fungal infection. DIAGNOSIS: A series of tests eventually confirmed the exclusive presence of HCoV-OC43 by the FilmArray Respiratory Panel from a throat swab sample. INTERVENTIONS: After the diagnosis was confirmed, the antimicrobial agents initially administered were discontinued. OUTCOMES: Although the chest CT scan images looked severe, the clinical course of the infection induced pneumonia was benign. The respiratory status of the patient was completely resolved in 2 weeks. LESSONS: This report highlights the importance of early identification of respiratory viruses, via the realization of their clinical characteristics, which helps reduce the duration of administration of antimicrobial agents in this setting.


Assuntos
Infecções por Coronavirus/diagnóstico , Pneumonia Viral/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras/complicações , Criança , Infecções por Coronavirus/complicações , Coronavirus Humano OC43/isolamento & purificação , Neutropenia Febril/complicações , Feminino , Humanos , Hospedeiro Imunocomprometido , Pneumonia Viral/complicações , Reação em Cadeia da Polimerase
10.
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi ; 32(4): 355-360, 2020 Aug 24.
Artigo em Chinês | MEDLINE | ID: mdl-32935508

RESUMO

OBJECTIVE: To investigate the biological properties of Schistosoma japonicum SjGrpE protein, and to express and purify the recombinant SjGrpE protein and test its immunogenicity. METHODS: The amino acid composition, molecular weight, hydrophilicity and hydrophobicity, transmembrane region, signal peptide, localization, phosphorylation site, ubiquitination site, glycosylation site, secondary and tertiary structures and B cell epitopes of the SjGrpE protein were predicted using bioinformatics analyses. The SjGrpE gene was amplified using PCR assay using S. japonicum cDNA as a template, double enzyme-digested and linked to the pET28a vector to yield the recombinant plasmid pET28a-SjGrpE. The recombinant plasmid pET28a-SjGrpE was transformed into Escherichia coli BL21, and then IPTG was employed to induce the expression of the target protein, which was purified by nickel ion affinity chromatography. After mice were immunized with the recombinant SjGrpE protein, mouse sera were collected, and the polyclonal antibody against the SjGrpE protein was characterized. RESULTS: SjGrpE protein, which was identified as a hydrophilic protein, was predicted to have a molecular weight of approximately 24.3 kDa without transmembrane regions or signal peptides, and locate in the mitochondrion. SjGrpE protein contained 18 phosphorylation sites and 2 ubiquitination sites, but had no glycosylation sites. In addition, SjGrpE protein contained 5 B-cell epitopes. The full length of SjGrpE gene was approximately 660 bp. The recombinant pET28a-SjGrpE plasmid was successfully generated, and the recombinant SjGrpE protein was obtained following the affinity chromatography, which stimulated mice to secrete high-titer antibodies. CONCLUSIONS: The recombinant SjGrpE protein has been successfully prepared and this recombinant protein has a high immunogenicity, which provides a basis for evaluating its value as a vaccine candidate for S. japonicum infections.


Assuntos
Proteínas de Helminto , Proteínas Recombinantes , Schistosoma japonicum , Animais , Anticorpos Anti-Helmínticos/isolamento & purificação , Clonagem Molecular , Escherichia coli/genética , Proteínas de Helminto/química , Proteínas de Helminto/genética , Proteínas de Helminto/imunologia , Proteínas de Helminto/isolamento & purificação , Camundongos , Plasmídeos/genética , Reação em Cadeia da Polimerase , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Schistosoma japonicum/genética , Schistosoma japonicum/metabolismo
11.
Zhongguo Zhong Yao Za Zhi ; 45(15): 3666-3671, 2020 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-32893556

RESUMO

Gastrodia elata is a kind of traditional Chinese medicinal materials and has good medicinal value. G. elata is divided into five varieties, which includes G. elata f. elata(proto variant), G. elata f. glauca, G. elata f. viridis, G. elata f. flavid and G. elata f. alba. Among them, G. elata f. elata and G. elata f. glauca have excellent characteristics and higher contents of gastrodin and polysaccharides. The hybrid of G. elata f. elata and G. elata f. glauca is present in markets, but the characteristics between hybrid and parent are not obvious and distinguished quickly and accurately. The aim of this study is to establish a PCR specific PCR identification method, which can identify G. elata f. elata, G. elata f. glauca and their hybrid. Based on the re-sequencing results of G. elata, we screened for the single nucleotide polymorphism(SNP) variation sites, and designed two pairs of specific primers(W291-F/W291-R and H255-F/H255-R). We further collected G. elata f. elata, G. elata f. glauca and their hybrid samples from different regions, established and optimized PCR method, and investigated and verified their tolerance and applicability. The results showed that when the annealing temperature was 48 ℃ and the number of cycles was 33, 255 bp specific band were obtained from G. elata f. glauca and hybrid by using specific primers W291-F/W291-R. When the annealing temperature was 51 ℃ and the number of cycles was 33, 291 bp specific band were obtained from G. elata f. elata and hybrid by using specific primers H255-F/H255-R. Our method could be used as a promising method to identify G. elata f. elata, G. elata f. glauca and their hybrid.


Assuntos
Gastrodia , Reação em Cadeia da Polimerase
12.
Mem Inst Oswaldo Cruz ; 115: e200220, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32935751

RESUMO

BACKGROUND: The Nyssomyia genus and Lutzomyia subgenus include medical important species that are Latin American leishmaniases vectors. Little is known about the phylogenetic relationships of closely-related species in each of these taxonomic groups that are morphologically indistinguishable or differentiated by very subtle details. OBJECTIVES: We inferred the phylogenetic relationships of closely-related species within both the Nyssomyia genus and the Lutzomyia subgenus using a cytochrome c oxidase subunit I (COI) fragment. METHODS: The sampling was carried out from 11 Argentinean localities. For genetic analyses, we used GenBank sequences in addition to our sequences from Argentina. Kimura 2-parameter (K2P) genetic distance and nucleotide divergence (Da) was calculated between closely-related species of Nyssomyia genus, Lutzomyia subgenus and between clades of Lutzomyia longipalpis complex. FINDINGS: The K2P and Da values within species of Nyssomyia genus and Lutzomyia subgenus were lower than the divergence detected between clades of Lu. longipalpis complex. The haplotype network analyses within Lutzomyia subgenus showed shared haplotypes between species, contrary to Nyssomyia genus with none haplotype shared. Bayesian inference within Nyssomyia genus presented structuring by species. MAIN CONCLUSIONS: This study evidences the phylogenetic proximity among closely-related species within Nyssomyia genus and Lutzomyia subgenus. The COI sequences of Nyssomyia neivai derived from the present study are the first available in GenBank.


Assuntos
Psychodidae/classificação , Psychodidae/genética , Animais , Argentina , Sequência de Bases , Teorema de Bayes , Leishmaniose , Filogenia , Reação em Cadeia da Polimerase/métodos , Análise de Sequência de DNA/métodos
13.
Clin Imaging ; 67: 219-225, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32871426

RESUMO

Since first report of a novel coronavirus in December of 2019, the Coronavirus Disease 2019 (COVID-19) pandemic has crippled healthcare systems around the world. While many initial screening protocols centered around laboratory detection of the virus, early testing assays were thought to be poorly sensitive in comparison to chest computed tomography, especially in asymptomatic disease. Coupled with shortages of reverse transcription polymerase chain reaction (RT-PCR) testing kits in many parts of the world, these regions instead turned to the use of advanced imaging as a first-line screening modality. However, in contrast to previous Severe Acute Respiratory Syndrome and Middle East Respiratory Syndrome coronavirus epidemics, chest X-ray has not demonstrated optimal sensitivity to be of much utility in first-line screening protocols. Though current national and international guidelines recommend for the use of RT-PCR as the primary screening tool for suspected cases of COVID-19, institutional and regional protocols must consider local availability of resources when issuing universal recommendations. Successful containment and social mitigation strategies worldwide have been thus far predicated on unified governmental responses, though the underlying ideologies of these practices may not be widely applicable in many Western nations. As the strain on the radiology workforce continues to mount, early results indicate a promising role for the use of machine-learning algorithms as risk stratification schema in the months to come.


Assuntos
Técnicas de Laboratório Clínico/métodos , Controle de Doenças Transmissíveis/métodos , Infecções por Coronavirus/diagnóstico , Saúde Global , Programas de Rastreamento/métodos , Pandemias , Pneumonia Viral/diagnóstico , Ásia , Betacoronavirus , Coronavirus , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/virologia , Europa (Continente) , Humanos , América do Norte , Pandemias/prevenção & controle , Pneumonia Viral/epidemiologia , Pneumonia Viral/prevenção & controle , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase/métodos , Estudos Retrospectivos , Tomografia Computadorizada por Raios X/métodos
14.
BMC Infect Dis ; 20(1): 659, 2020 Sep 07.
Artigo em Inglês | MEDLINE | ID: mdl-32894092

RESUMO

BACKGROUND: Diarrheagenic Escherichia coli (DEC) are among the leading pathogens associated with endemic diarrhea in low income countries. Yet, few epidemiological studies have focused the contribution of enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli (EIEC) and diffusely adherent E. coli (DAEC). METHODS: We assessed the contribution of EHEC, EIEC and DAEC isolated from stool samples from a case-control study conducted in children aged < 5 years in Southern Mozambique between December 2007 and November 2012. The isolates were screened by conventional PCR targeting stx1 and stx2 (EHEC), ial and ipaH (EIEC), and daaE (DAEC) genes. RESULTS: We analyzed 297 samples from cases with less-severe diarrhea (LSD) matched to 297 controls, and 89 samples from cases with moderate-to-severe diarrhea (MSD) matched to 222 controls, collected between November 3, 2011 and November 2, 2012. DEC were more common among LSD cases (2.7%, [8/297] of cases vs. 1.3% [4/297] of controls; p = 0.243]) than in MSD cases (0%, [0/89] of cases vs. 0.4%, [1/222] of controls; p = 1.000). Detailed analysis revealed low frequency of EHEC, DAEC or EIEC and no association with diarrhea in all age strata. Although the low frequency, EIEC was predominant in LSD cases aged 24-59 months (4.1% for cases vs. 0% for controls), followed by DAEC in similar frequency for cases and controls in infants (1.9%) and lastly EHEC from one control. Analysis of a subset of samples from previous period (December 10, 2007 and October 31, 2011) showed high frequency of DEC in controls compared to MSD cases (16.2%, [25/154] vs. 11.9%, [14/118], p = 0.383, respectively). Among these, DAEC predominated, being detected in 7.7% of cases vs. 17.6% of controls aged 24-59 months, followed by EIEC in 7.7% of cases vs. 5.9% of controls for the same age category, although no association was observed. EHEC was detected in one sample from cases and two from controls. CONCLUSIONS: Our data suggests that although EHEC, DAEC and EIEC are less frequent in endemic diarrhea in rural Mozambique, attention should be given to their transmission dynamics (e.g. the role on sporadic or epidemic diarrhea) considering that the role of asymptomatic individuals as source of dissemination remains unknown.


Assuntos
Diarreia/epidemiologia , Escherichia coli Êntero-Hemorrágica/genética , Infecções por Escherichia coli/epidemiologia , Saúde da População Rural , Estudos de Casos e Controles , Pré-Escolar , Diarreia/microbiologia , Doenças Endêmicas , Escherichia coli Êntero-Hemorrágica/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Fezes/microbiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Moçambique/epidemiologia , Reação em Cadeia da Polimerase , Prevalência , População Rural
16.
Plast Reconstr Surg ; 146(2): 248e-250e, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32740631
17.
BMC Infect Dis ; 20(1): 587, 2020 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-32770954

RESUMO

BACKGROUND: Tuberculosis (TB) is transmitted in bioaerosols containing Mycobacterium tuberculosis (Mtb). Despite being central to ongoing TB transmission, no routine diagnostic assay exists to measure Mtb in bioaerosols. Furthermore, published studies of Mtb in bioaerosol samples have been limited to individuals with sputum-positive pulmonary TB. Notably, TB diagnosis is based on clinical symptoms and sputum laboratory findings. This is despite the fact that approximately half of all patients commencing TB treatment are sputum-negative, resulting in a high proportion of presumptive treatments. Here, we propose to use a sensitive air sampling protocol to investigate the prevalence of Mtb-containing bioaerosols in both sputum-positive and sputum-negative TB suspects, at the same time evaluating the potential to identify unrecognized transmitters of TB. METHODS: Our parallel-group design will identify viable Mtb in bioaerosols produced by individuals attending a TB clinic in South Africa. Sampling will be performed on eligible individuals presenting with symptoms indicative of TB and repeated at 14 days if initially positive. Participants will be prospectively classified into three distinct groups based on National TB Control Program (NTBCP) criteria: Group A, TB notification with sputum-based laboratory confirmation; Group B, TB notification with empiric diagnosis; and Group C, individuals not notified. Group C individuals with detectable Mtb bioaerosol will be monitored until resolution of clinical and laboratory status. Collection of bioaerosol specimens will be via two consecutive sampling modalities: (1) direct sampling following a specific respiratory manoeuvre; and (2) indirect sampling during passive respiratory activity. Bioaerosol specimens will be analyzed for viable Mtb using DMN-trehalose staining and live-cell fluorescence microscopy. Mtb genomes and mycobacterial and host lipids will be detected using droplet digital PCR and mass spectrometry analyses, respectively. The primary objective is to determine the prevalence of Mtb bioaerosols in all TB clinic attendees and in each of the groups. Secondary objectives are to investigate differences in prevalence of Mtb bioaerosol by HIV status and current isoniazid preventive therapy (IPT) use; we will also determine the impact of anti-TB chemotherapy on Mtb-containing bioaerosol production. DISCUSSION: Respiratory bioaerosol has a potential role in non-invasive TB diagnosis, infectivity measurement and treatment monitoring. TRIAL REGISTRATION: ClinicalTrials.gov: NCT04241809 . Date of Registration: 27/1/2020.


Assuntos
Aerossóis/análise , Manejo de Espécimes/métodos , Tuberculose Pulmonar/diagnóstico , Adulto , DNA Bacteriano/química , DNA Bacteriano/metabolismo , Feminino , Humanos , Masculino , Espectrometria de Massas , Pessoa de Meia-Idade , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Reação em Cadeia da Polimerase , África do Sul , Escarro/microbiologia
18.
PLoS One ; 15(8): e0237960, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32822413

RESUMO

BACKGROUND: In addition to the lack of COVID-19 diagnostic tests for the whole Spanish population, the current strategy is to identify the disease early to limit contagion in the community. AIM: To determine clinical factors of a poor prognosis in patients with COVID-19 infection. DESIGN AND SETTING: Descriptive, observational, retrospective study in three primary healthcare centres with an assigned population of 100,000. METHOD: Examination of the medical records of patients with COVID-19 infections confirmed by polymerase chain reaction. Logistic multivariate regression models adjusted for age and sex were constructed to analyse independent predictive factors associated with death, ICU admission and hospitalization. RESULTS: We included 322 patients (mean age 56.7 years, 50% female, 115 (35.7%) aged ≥ 65 years): 123 (38.2) were health workers (doctors, nurses, auxiliaries). Predictors of ICU admission or death were greater age (OR = 1.05; 95%CI = 1.03 to 1.07), male sex (OR = 2.94; 95%CI = 1.55 to 5.82), autoimmune disease (OR = 2.82; 95%CI = 1.00 to 7.84), bilateral pulmonary infiltrates (OR = 2.86; 95%CI = 1.41 to 6.13), elevated lactate-dehydrogenase (OR = 2.85; 95%CI = 1.28 to 6.90), elevated D-dimer (OR = 2.85; 95%CI = 1.22 to 6.98) and elevated C-reactive protein (OR = 2.38; 95%CI = 1.22 to 4.68). Myalgia or arthralgia (OR = 0.31; 95%CI = 0.12 to 0.70) was protective factor against ICU admission and death. Predictors of hospitalization were chills (OR = 5.66; 95%CI = 1.68 to 23.49), fever (OR = 3.33; 95%CI = 1.89 to 5.96), dyspnoea (OR = 2.92; 95%CI = 1.62 to 5.42), depression (OR = 6.06; 95%CI = 1.54 to 40.42), lymphopenia (OR = 3.48; 95%CI = 1.67 to 7.40) and elevated C-reactive protein (OR = 3.27; 95%CI = 1.59 to 7.18). Anosmia (OR = 0.42; 95%CI = 0.19 to 0.90) was the only significant protective factor for hospitalization after adjusting for age and sex. CONCLUSION: Determining the clinical, biological and radiological characteristics of patients with suspected COVID-19 infection will be key to early treatment and isolation and the tracing of contacts.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Adolescente , Adulto , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Infecções por Coronavirus/mortalidade , Infecções por Coronavirus/virologia , Feminino , Hospitalização , Humanos , Unidades de Terapia Intensiva , Masculino , Pessoa de Meia-Idade , Mortalidade , Pandemias , Pneumonia Viral/mortalidade , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase , Prognóstico , Fatores de Proteção , Estudos Retrospectivos , Fatores de Risco , Fatores Sexuais , Espanha/epidemiologia , Adulto Jovem
19.
Genes (Basel) ; 11(8)2020 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-32806776

RESUMO

Deep knowledge of the genetic features of SARS-CoV-2 is essential to track the ongoing pandemic through different geographical areas and to design and develop early diagnostic procedures, therapeutic strategies, public health interventions, and vaccines. We describe protocols and first results of the Ion AmpliSeq™ SARS-CoV-2 Research Panel by a massively parallel sequencing (MPS) assay. The panel allows for targeted sequencing by overlapping amplicons, thereby providing specific, accurate, and high throughput analysis. A modified reverse transcription reaction, which consists of the use of a SARS-CoV-2 specific primers pool from the Ion AmpliSeq SARS-CoV-2 Research Panel, was assessed in order to promote viral RNA specific reverse transcription. The aim of this study was to evaluate the effectiveness of the Ion AmpliSeq™ SARS-CoV-2 Research Panel in sequencing the entire viral genome in different samples. SARS-CoV-2 sequence data were obtained from ten viral isolates and one nasopharyngeal swab from different patients. The ten isolate samples amplified with 12 PCR cycles displayed high mean depth values compared to those of the two isolates amplified with 20 PCR cycles. High mean depth values were also obtained for the nasopharyngeal swab processed by use of a target-specific reverse transcription. The relative depth of coverage (rDoC) analysis showed that when 12 PCR cycles were used, all target regions were amplified with high sequencing coverage, while in libraries amplified at 20 cycles, a poor uniformity of amplification, with absent or low coverage of many target regions, was observed. Our results show that the Ion AmpliSeq SARS-CoV-2 Research Panel can achieve rapid and high throughput SARS-CoV-2 whole genome sequencing from 10 ng of DNA-free viral RNA from isolates and from 1 ng of DNA-free viral RNA from a nasopharyngeal swab using 12 PCR cycles for library amplification. The modified RT-PCR protocol yielded superior results on the nasopharyngeal swab compared to the reverse transcription reaction set up according to the manufacturer's instructions.


Assuntos
Betacoronavirus/genética , Infecções por Coronavirus/virologia , Pneumonia Viral/virologia , Reação em Cadeia da Polimerase/métodos , Sequenciamento Completo do Genoma/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Betacoronavirus/patogenicidade , Chlorocebus aethiops , Primers do DNA/normas , Feminino , Genoma Viral , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Reação em Cadeia da Polimerase/normas , Células Vero , Sequenciamento Completo do Genoma/normas
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA