RESUMO
SARS-CoV-2 has recently emerged, becoming a global threat, affecting directly all human beings owing to its morbidity and mortality and indirectly, due to the enormous economic and psychological impact produced by social isolation, the most effective measure so far, but unsustainable for a long period. The scientific effort to understand and control SARS-CoV-2 transmission and clinical impact has been huge, and important achievements are highlighted in this review. Diagnosis is central and is the first step in recognizing and fighting any infectious agent. Instrumental to that is the quality of the data, relying on serological and molecular surveys in addition to trustworthy clinical records. However, the fast spread of a virus adapted for human-to-human respiratory transmission raised a demand for millions of molecular tests that are simply not available. Several candidate drugs are under evaluation in clinical trials. Those with an already recognized safety profile are more auspicious, since, if proven effective, can cut several steps of production and phase 2 and 3 trials. More than one hundred vaccine prototypes are in different stages of development, however, safety and efficacy evaluations cannot be obviated, implicating, most optimistically, in at least months for us to have an effective immunization, the definite measure to allow a safe return to the pre-pandemic lifestyle. Science has never been more necessary and present in daily life. Relying on the best of human wit is the only way out to this pandemic, saving as many lives as possible.
Assuntos
Infecções por Coronavirus , Pandemias , Pneumonia Viral , Antivirais/farmacologia , Betacoronavirus , COVID-19 , Vacinas contra COVID-19 , Ensaios Clínicos como Assunto , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/epidemiologia , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/terapia , Infecções por Coronavirus/transmissão , Avaliação de Medicamentos , Humanos , Pneumonia Viral/diagnóstico , Pneumonia Viral/epidemiologia , Pneumonia Viral/terapia , Pneumonia Viral/transmissão , SARS-CoV-2 , Vacinas ViraisRESUMO
SARS-CoV-2 has recently emerged, becoming a global threat, affecting directly all human beings owing to its morbidity and mortality and indirectly, due to the enormous economic and psychological impact produced by social isolation, the most effective measure so far, but unsustainable for a long period. The scientific effort to understand and control SARS-CoV-2 transmission and clinical impact has been huge, and important achievements are highlighted in this review. Diagnosis is central and is the first step in recognizing and fighting any infectious agent. Instrumental to that is the quality of the data, relying on serological and molecular surveys in addition to trustworthy clinical records. However, the fast spread of a virus adapted for human-to-human respiratory transmission raised a demand for millions of molecular tests that are simply not available. Several candidate drugs are under evaluation in clinical trials. Those with an already recognized safety profile are more auspicious, since, if proven effective, can cut several steps of production and phase 2 and 3 trials. More than one hundred vaccine prototypes are in different stages of development, however, safety and efficacy evaluations cannot be obviated, implicating, most optimistically, in at least months for us to have an effective immunization, the definite measure to allow a safe return to the pre-pandemic lifestyle. Science has never been more necessary and present in daily life. Relying on the best of human wit is the only way out to this pandemic, saving as many lives as possible.
Assuntos
Humanos , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/terapia , Infecções por Coronavirus/transmissão , Infecções por Coronavirus/epidemiologia , Pandemias , Antivirais/farmacologia , Pneumonia Viral/diagnóstico , Pneumonia Viral/terapia , Pneumonia Viral/epidemiologia , Vacinas Virais , Ensaios Clínicos como Assunto , Avaliação de Medicamentos , Betacoronavirus , Vacinas contra COVID-19 , COVID-19RESUMO
OBJECTIVE: The aim of our work was to establish a semi-automated high-throughput DNA amplification method for the universal screening of bacteria in platelet concentrates (PCs). BACKGROUND: Among cases of transfusion transmission of infectious agents, bacterial contamination ranks first in the number of events, morbidity and mortality. Transmission occurs mainly by transfused PCs. Automated culture is adopted by some blood banks for screening of bacterial contamination, but this procedure is expensive and has a relatively long turnaround time. METHODS: PCs were spiked with suspensions of five different bacterial species in a final concentration of 1 and 10 colony-forming units (CFU) per millilitre. After incubation, the presence of bacteria was investigated by real-time polymerase chain reaction (PCR) and by the Enhanced Bacterial Detection System (eBDS, Pall) assay as a reference method. Real-time PCR amplification was performed with a set of universal primers and probes targeting the 16S rRNA gene. Co-amplification of human mitochondrial DNA served as an internal control. RESULTS: Using the real-time PCR method, it was possible to detect the presence of all bacterial species tested with an initial concentration of 10 CFU mL-1 24 h after contamination, except for Staphylococcus hominis. The PCR assay also detected, at 24 h, the presence of Serratia marcescens and Enterobacter cloacae with an initial concentration of 1 CFU mL-1 . CONCLUSIONS: The real-time PCR assay may be a reliable alternative to conventional culture methods in the screening of bacterial contamination of PCs, enabling bacterial detection even with a low initial concentration of microorganisms.
Assuntos
Bactérias/genética , Doadores de Sangue , Plaquetas/microbiologia , Genes de RNAr/genética , Técnicas de Amplificação de Ácido Nucleico , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Brasil , HumanosRESUMO
Persistent human papillomavirus (HPV) infection is an essential factor of cervical cancer. This study evaluated the analytical performance of restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) assay compared to PapilloCheck® microarray to identify human papilloma virus (HPV) in cervical cells. Three hundred and twenty-five women were analyzed. One sample was used for conventional cytology and another sample was collected using BD SurePath™ kit for HPV tests. Eighty samples (24.6%) were positive for HPV gene by PCR-Multiplex and were then submitted to PCR-RFLP and PapilloCheck® microarray. There was a genotyping agreement in 71.25% (57/80) on at least one HPV type between PCR-RFLP and PapilloCheck® microarray. In 22 samples (27.5%), the results were discordant and those samples were additionally analyzed by DNA sequencing. HPV 16 was the most prevalent HPV type found in both methods, followed by HPVs 53, 68, 18, 39, and 66 using PCR-RFLP analysis, and HPVs 39, 53, 68, 56, 31, and 66 using PapilloCheck® microarray. In the present study, a perfect agreement using Cohen's kappa (κ) was found in HPV 33 and 58 (κ=1), very good for HPV 51, and good for types 16, 18, 53, 59, 66, 68, 70, and 73. PCR-RFLP analysis identified only 25% (20/80) HPV coinfection, and PapilloCheck® microarray found 62.5% (50/80). Our Cohen's kappa results indicate that our in-house HPV genotyping testing (PCR-RFLP analysis) could be applied as a primary HPV test screening, especially in low income countries. If multiple HPV types are found in this primary test, a more descriptive test, such as PapilloCheck® microarray, could be performed.
Assuntos
Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adolescente , Adulto , Idoso , Detecção Precoce de Câncer , Feminino , Genótipo , Humanos , Programas de Rastreamento , Análise em Microsséries , Pessoa de Meia-Idade , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sensibilidade e Especificidade , Neoplasias do Colo do Útero/virologia , Adulto JovemRESUMO
Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) molecule is expressed on T-lymphocyte membrane and negatively influences the antigen-presenting process. Reduced expression of CTLA-4 due to gene polymorphisms is associated with increased risk of autoimmune disorders, whose physiopathology is similar to that of post-transfusion red blood cell (RBC) alloimmunization. Our goal was to evaluate if polymorphisms of CTLA-4 gene that affect protein expression are associated with RBC alloimmunization. This was a case-control study in which 134 sickle cell disease (SCD) patients and 253 non-SCD patients were included. All patients were genotyped for the polymorphisms 49A/G and -318C/T of CTLA-4 gene. The genotype frequency of -318C/T differed significantly between alloimmunized and nonalloimmunized SCD patients, irrespective of clinical confounders (p = .016). SCD patients heterozygous for -318T allele presented higher risk of alloantibody development (OR: 5.4, CI: 1.15-25.6). In conclusion, the polymorphism -318C/T of CTLA-4 gene is associated with RBC alloimmunization among SCD patients. This highlights the role played by CTLA-4 on post-transfusion alloantibody development.
Assuntos
Anemia Falciforme/genética , Doenças Autoimunes/genética , Antígeno CTLA-4/genética , Eritrócitos/imunologia , Adulto , Anemia Falciforme/sangue , Anemia Falciforme/imunologia , Anemia Falciforme/prevenção & controle , Células Apresentadoras de Antígenos/imunologia , Células Apresentadoras de Antígenos/patologia , Doenças Autoimunes/sangue , Doenças Autoimunes/imunologia , Doenças Autoimunes/patologia , Antígeno CTLA-4/imunologia , Eritrócitos/patologia , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Imunização/efeitos adversos , Isoanticorpos/imunologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único/genética , Fatores de Risco , Linfócitos T/imunologia , Linfócitos T/patologiaRESUMO
BACKGROUND: Blood donors are, in principle, healthy individuals who may be revealed as infectious for blood-borne agents by the laboratory screening process, depicting the asymptomatic burden of the disease. Therefore, monitoring hepatitis C virus (HCV)-infected donor and human immunodeficiency virus (HIV)-infected donor and associating to their demographical and behavioural characteristics may shed light on the dynamics and contemporary changes in these viruses' epidemiology. METHODS: Donors presenting repeatedly reactive HCV or HIV serology/nucleic acid testing (NAT) screening results were submitted to confirmatory testing. Confirmed positive donors were invited to return to the blood bank for notification and counselling when a follow-up sample was obtained and an interview performed to eventually disclose potential risks. HCV- or HIV-infected donors identified over 11 years of screening (2004-2015) were evaluated for demographic and behavioural parameters. RESULTS: In the period, 139 160 donations were screened, and 36 (0.025%) were found positive for HIV, stemming from 29 male and 7 female donors. Among those, eight subjects were repeat donors. A total of 95 donations were found repeatedly reactive for HCV (0.068%), obtained from 60 men and 35 women. Noticeably, in despite of a higher HCV prevalence in the donor population, the incidence of HIV among repeat donors was 10 times that of HCV (18 × 1.6/100 000 persons-year, respectively). On average, HIV-seroreactive men were found to be younger (mean = 34 years old) than women (mean = 40 years old). A total of 10 donors acknowledged sexual behaviours not previously informed, including 2 who were aware of their HIV-positive status and another 2 who admitted to be seeking HIV testing. No window period donation was verified. DISCUSSION: The majority of the HIV-infected donors are young males who deny risk factors in the interview and also ignore the confidence self-exclusion opportunity. As they may reiterate this behaviour in serial donations, use of the most sensitive laboratory testing is justified in this setting.
Assuntos
Doadores de Sangue , Seleção do Doador/métodos , Infecções por HIV , Hepatite C , Técnicas de Amplificação de Ácido Nucleico , RNA Viral/sangue , Adulto , Brasil/epidemiologia , Feminino , Infecções por HIV/sangue , Infecções por HIV/epidemiologia , Hepatite C/sangue , Hepatite C/epidemiologia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos SoroepidemiológicosRESUMO
The association between high-risk human papillomavirus (HPV) genotypes and p16 expression in indigenous women from the Xingu Indigenous Park, Brazil, was unknown. This study evaluated p16 expression in women with a histological diagnosis of cervical intraepithelial neoplasia (CIN) 3 or higher and correlated this expression with HPV genotypes to determine possible discrepancies in the expression of this marker. We evaluated 37 previously collected samples with different HPV genotypes and high-grade lesions diagnosed based on cytology, histology, and colposcopy. Immunohistochemical analysis was performed using paraffin-embedded tissue sections and the CINtec® Histology Kit. p16 protein expression was investigated by immunostaining with an anti-p16 antibody. HPV genotyping was performed by reverse hybridization. The age of the study population ranged from 22-75 years (43.81 ± 15.89 years) and parity ranged from 1-11 (5.92 ± 2.58). Thirteen different HPV genotypes were found using the INNO-LiPA kit. Single and multiple infections by HPV were found with prevalence of single infections (P = 0.029). Comparison between HPV genotype and simple or multiple infections was highly significant; it was observed more HPV 52 followed by HPV 16 in single infections (P < 0.001). p16 expression was predominantly diffuse, which was observed in 91.7% of lesions, whereas 8.3% were focal (P < 0.001). HPV 52, HPV 16 and 31 were the most prevalent HPV types in high-grade CIN in these indigenous women. Diffuse p16 expression in high-grade CIN was not influenced by the viral genotype; however, more studies are necessary to further our understanding of this restricted group.
Assuntos
Biomarcadores Tumorais/biossíntese , Inibidor p16 de Quinase Dependente de Ciclina/biossíntese , Infecções por Papillomavirus/genética , Displasia do Colo do Útero/genética , Adulto , Idoso , Biomarcadores Tumorais/genética , Brasil , Colposcopia , Inibidor p16 de Quinase Dependente de Ciclina/genética , Feminino , Regulação da Expressão Gênica , Genótipo , Papillomavirus Humano 16/genética , Papillomavirus Humano 16/isolamento & purificação , Papillomavirus Humano 16/patogenicidade , Papillomavirus Humano 31/genética , Papillomavirus Humano 31/isolamento & purificação , Papillomavirus Humano 31/patogenicidade , Humanos , Pessoa de Meia-Idade , Infecções por Papillomavirus/patologia , Infecções por Papillomavirus/virologia , Displasia do Colo do Útero/patologia , Displasia do Colo do Útero/virologiaRESUMO
BACKGROUND AND OBJECTIVES: Serological screening for the Vel- phenotype is complex given the large individual variation in the levels of expression of the Vel antigen, and the polyclonal anti-human sera of immunised persons, when available, show heterogeneous reactivity levels. Studies of the SMIM1 gene have enabled the development of several molecular methodologies that will be crucially important for the screening of different populations, including Brazilians. To evaluate the deletion of 17 bp in the SMIM1 gene in a population from the south of Brazil, 448 unrelated blood donors from 7 regions comprising the haemotherapy network in the state of Santa Catarina were evaluated between August 2011 and March 2014. MATERIALS AND METHODS: DNA samples from these donors were analysed employing a 5' nuclease real-time polymerase chain reaction (PCR) assay targeting the 17 bp deletion in the SMIM1 gene. RESULTS: Among the 448 samples analysed, 10 (2·23%) harboured the 17 bp deletion of the gene SMIM1, and all were heterozygote for the SMIM1*64_80 del allele. CONCLUSION: The allelic frequency found differed from those observed in other Caucasian populations. This difference can be explained by the ethnic make-up of each Caucasian population. The data obtained are important to characterise the correct phenotype of the donor as the serological assay results are not reliable due to variations in the expression intensity of the Vel antigen in heterozygote donors for the SMIM1*64_80 del allele. Moreover, the tool used in this study is of great value for identifying a donor Vel- phenotype and supplying a possible need for transfusion.
Assuntos
Alelos , Sequência de Bases , Doadores de Sangue , Proteínas de Membrana/genética , Polimorfismo de Fragmento de Restrição , Deleção de Sequência , Brasil , Feminino , Humanos , Masculino , Reação em Cadeia da Polimerase em Tempo RealRESUMO
HIV and human papillomavirus (HPV) coinfection is increasing, especially in the anal canal (AC) and cervico-vaginal regions. We identified anal epithelium abnormalities related to high-risk HPV (HR-HPV) lesions in the lower genital tracts (LGTs) of HIV-positive women, described the HPV genotypes identified, and assessed the expression of E6/E7 oncogenes in coinfected patients. Ninety-eight women were enrolled in groups combining HIV status and presence or absence of HPV in the LGT. Anal and cervical smears were collected for cytology and HR-HPV assays using Cobas(®) and/or PapilloCheck(®). Samples with highly oncogenic HPV genotypes were confirmed by NucliSENS EasyQ(®). Forty-two HIV-positive (25-52; mean age 39.5) and 56 HIV-negative (18-58; mean age 35.7) patients were included. E2 and C1 groups presented AC alterations (P = 0.002); altered images for high-resolution anoscopy were higher in E1 and C2 (P < 0.001). Of the 29 women with alterations, 41.38% were HIV-negative and 58.62% were HIV-positive (P < 0.001). HIV-positive patients accounted for 29% of the anal high-grade squamous intraepithelial lesions (P = 0.015). The Cobas(®) positive result frequency was higher in three AC groups than in the other groups. There was variation in the number of HPV types in the cervico-vaginal samples among the study groups (P < 0.001). Anal cytology and anoscopy showed more altered findings in HIV-positive patients with HPV in the LGT. HR-HPV anal infections by various genotypes are common and are associated with cervical infections in HIV-positive patients. E6/E7 expression is apparently more common in the AC of HIV-positive women.
Assuntos
Coinfecção/virologia , Infecções por HIV/virologia , Infecções por Papillomavirus/virologia , Infecções do Sistema Genital/virologia , Adolescente , Adulto , Canal Anal/patologia , Canal Anal/virologia , Coinfecção/complicações , Coinfecção/patologia , Feminino , Genótipo , HIV/genética , HIV/isolamento & purificação , HIV/patogenicidade , Infecções por HIV/complicações , Infecções por HIV/genética , Infecções por HIV/patologia , Humanos , Pessoa de Meia-Idade , Proteínas Oncogênicas Virais/biossíntese , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Papillomaviridae/patogenicidade , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/patologia , RNA Mensageiro/biossíntese , Proteínas Repressoras/biossíntese , Infecções do Sistema Genital/complicações , Infecções do Sistema Genital/genética , Infecções do Sistema Genital/patologia , Esfregaço VaginalRESUMO
OBJECTIVE: To identify the demographic characteristics, risk factors and motivations for donating among blood donors with reactive serologic tests for syphilis. BACKGROUND: Post-donation interviews with syphilis seropositive blood donors improve recruitment and screening strategies. METHODS: This case-control study compares 75 Venereal Disease Research Laboratory (VDRL) > 8, EIA+ (enzyme immunoassay) and FTA-ABS+ (fluorescent treponemal antibody); 80 VDRL-, EIA+ and FTA-ABS+; and 34 VDRL- and EIA- donors between 2004 and 2009. Donors were assessed by their demographic characteristics, sexual behaviour, history of alcohol and illicit drugs use, and motivations to donate. RESULTS: Donors with VDRL > 8 were more likely to be divorced [AOR = 12·53; 95% confidence interval (CI) 1·30-120·81], to have had more than six sexual partners (AOR=7·1; 95% CI 1·12-44·62) and to report male-male-sex in the past 12 months (AOR=8·18; 95% CI 1·78-37·60). Donors with VDRL-, EIA+ and FTA-ABS+ were less likely to be female (AOR=0·26; 95% CI 0·07-0·96), more likely to be older (AOR=10·2; 95% CI 2·45-42·58 ≥ 39 and <60 years old) and to have had more than six sexual partners in the past 12 months (AOR = 8·37; 95% CI 1·49-46·91). There was no significant difference among groups regarding illicit drugs use; 30·7% (VDRL > 8) and 12·5% (VDRL-, EIA+ and FTA-ABS+) of donors reported that they had been at risk for HIV infection (P = 0·004). One-third of donors came to the blood bank to help a friend or a relative who needed blood. CONCLUSION: Although donors exposed to syphilis reported and recognised some high risk behaviour, most were motivated by direct appeal to donate blood. Monitoring the risk profile of blood donors can benefit public health and improve blood safety.