RESUMO
Today, PCR using broad-range primers is being used increasingly to detect pathogens from resected heart valves. Herein is described the first case of multivalve infective endocarditis where 16S rDNA PCR was used to detect a single pathogen from two affected valves in a 61-year-old man. Triple heart valve replacement was required despite six weeks of appropriate antimicrobial therapy. The organism was confirmed as Streptococcus gallolyticus subsp. macedonicus, a member of the 'S. equinus/S. bovis' complex. To date, only one report has been made of human infection due to this organism. This may be due to the limited resolution of the routine diagnostic methods used and/or as a consequence of the complex nomenclature associated with this group of organisms.
Assuntos
Técnicas Bacteriológicas , DNA Ribossômico/genética , Endocardite Bacteriana/microbiologia , Reação em Cadeia da Polimerase , Infecções Estreptocócicas/microbiologia , Streptococcus/genética , Endocardite Bacteriana/cirurgia , Doenças das Valvas Cardíacas/microbiologia , Doenças das Valvas Cardíacas/cirurgia , Implante de Prótese de Valva Cardíaca , Valvas Cardíacas/microbiologia , Valvas Cardíacas/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Infecções Estreptocócicas/cirurgia , Streptococcus bovis/genética , Streptococcus equi/genéticaRESUMO
PURPOSE: Cervical cancer is the most frequently diagnosed female cancer in The Gambia, representing approximately 30â% of cases. In 2014, the quadrivalent human papilloma virus (HPV) vaccine was introduced, which offers protection against HPV genotypes 6, 11, 16 and 18. To evaluate the potential effectiveness of this vaccine, genotype distribution and risk factor analysis were assessed. METHODOLOGY: Endocervical samples (n=232) were collected from women aged 20-49 years residing in urban Gambia. A questionnaire was administered to capture socio-demographic and cervical cancer risk factors. HPV detection and genotyping was performed by PCR amplification of the L1 major capsid gene and analysis of sequenced PCR products.Results/Key findings. The prevalence of HPV was 12â% (28/232), and the high-risk (HR) genotype HPV 52 (5/28) was the most prevalent genotype. HR-HPV sequences had high identity (≥90â%) to isolates which originated from America, Europe and Asia but not from Africa. Half (14/28) of participants were co-infected with Ureaplasma urealyticum/parvum, which increases the risk of progression to cervical cancer. Female genital mutilation and the use of hormone contraception for >5 years were identified as potential risk factors for HPV infection. Ethnicity-associated differences were also noted; participants of the Fula ethnic group had a higher prevalence of HR-HPV infection (31.3â%) compared to the Mandinka (18.8â%) and Wollof (12.5â%) groups. CONCLUSION: These data may have a significant public health impact as the HPV quadrivalent vaccine may be of limited value if the circulating non-HPV 16/18 HR-genotypes are responsible for cytological abnormalities of the cervix.
Assuntos
Genótipo , Papillomaviridae/genética , Infecções por Papillomavirus/epidemiologia , Displasia do Colo do Útero/virologia , Neoplasias do Colo do Útero/virologia , Adulto , Proteínas do Capsídeo/genética , Coinfecção/epidemiologia , Coinfecção/microbiologia , Coinfecção/virologia , Anticoncepção/efeitos adversos , Análise Fatorial , Feminino , Gâmbia/epidemiologia , Genitália Feminina/lesões , Humanos , Pessoa de Meia-Idade , Proteínas Oncogênicas Virais/genética , Papillomaviridae/classificação , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/microbiologia , Infecções por Papillomavirus/virologia , Vacinas contra Papillomavirus/administração & dosagem , Vacinas contra Papillomavirus/efeitos adversos , Reação em Cadeia da Polimerase , Prevalência , Fatores de Risco , Inquéritos e Questionários , População Urbana , Infecções por Ureaplasma/epidemiologia , Infecções por Ureaplasma/microbiologia , Infecções por Ureaplasma/virologia , Ureaplasma urealyticum/isolamento & purificação , Neoplasias do Colo do Útero/epidemiologia , Potência de Vacina , Adulto Jovem , Displasia do Colo do Útero/epidemiologiaRESUMO
The limitations of classical diagnostic methods for invasive fungal infections (IFIs) have led to the development of molecular techniques to aid in the detection of IFIs. Despite good published performance, interlaboratory reproduction of these assays is variable, and no consensus has been reached for an optimal method. This publication describes the first multicenter study of polymerase chain reaction methods, for the detection of Aspergillus and Candida species, currently used in the UK and Ireland by distribution and analysis of multiple specimen control panels. All three Candida methods were comparable, achieving a satisfactory level of detection (10 cfu), and the method of preference was dependent on the requirements of the particular laboratory. The results for the five Aspergillus assays were more variable, but two methods (2Asp and 4Asp) were superior (10(1) conidia). Formally, the overall performances of the two Aspergillus assays were comparable (kappa statistic = 0.77). However, on the Roche LightCycler, there was a clear sample-type effect that greatly reduced the detection limit of the 4Asp method when testing whole blood samples. Therefore, the preferred Aspergillus method relied on the amplification platform available to the user. This study represents the initial process to achieve a consensus method for the diagnosis of IFIs.
Assuntos
DNA Fúngico/análise , Técnicas de Diagnóstico Molecular/métodos , Micoses/diagnóstico , Reação em Cadeia da Polimerase/métodos , Aspergilose/diagnóstico , Sequência de Bases , Candidíase/diagnóstico , Consenso , Humanos , Irlanda , Técnicas de Diagnóstico Molecular/instrumentação , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/instrumentação , Sensibilidade e Especificidade , Homologia de Sequência do Ácido Nucleico , Reino UnidoRESUMO
We have developed a novel real-time PCR assay to identify and perform preliminary genotyping of mycobacteria in a manner tailored to our local service. Within a single thermocycler run, mycobacterial 16S rDNA and the Mycobacterium tuberculosis global lineage-defining RD750 polymorphism are targeted in separate reaction tubes, each of which includes both TaqMan and SYBR Green chemistries. The results of this 16S-RD assay differentiate M. tuberculosis complex (MTBC) from nontuberculous mycobacteria (NTM) and recognize whether or not MTBC isolates belong to the East African-Indian lineage, the single most frequently isolated global MTBC lineage in our service. If required, NTM amplicons may be sequenced to provide more specific identities. We report the technical performance of this assay on 88 mycobacteria-positive cultures and discuss its use in the initial management of mycobacterial infections. The 16S-RD assay correctly identified all 70 MTBC-positive cultures and 17 NTM-positive cultures while contemporaneously recognizing 26 MTBC isolates as within and 44 outside the East African-Indian lineage. In artificial samples, the combined assay also showed limited potential to detect mixed mycobacterial infections (MTBC/NTM) and tuberculosis infections involving more than one global MTBC lineage. The approach we have established can be readily tailored to targets of particular value for any mycobacterial diagnostic service, thereby optimizing the value of the results for local clinical and public health management of mycobacterial infections.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Corantes Fluorescentes/metabolismo , Mycobacterium tuberculosis , Compostos Orgânicos/metabolismo , Reação em Cadeia da Polimerase/métodos , Polimorfismo Genético , Taq Polimerase/metabolismo , Técnicas de Tipagem Bacteriana/instrumentação , Benzotiazóis , DNA Bacteriano/análise , DNA Bacteriano/genética , Diaminas , Humanos , Mycobacterium tuberculosis/classificação , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Filogenia , Reação em Cadeia da Polimerase/instrumentação , Quinolinas , RNA Ribossômico 16S/análise , RNA Ribossômico 16S/genéticaRESUMO
Sexually transmitted infections (STIs) are an increasingly common and important cause of a fever in a returning traveler. Systemic complications of STIs, human immunodeficiency virus seroconversion illness, and secondary syphilis are diagnoses that can easily be missed. We present a case of culture-negative disseminated gonococcal infection presenting with fever, malaise, polyarthralgia, arthritis, and a rash that developed following orogenital contact and was diagnosed using real-time polymerase chain reaction. This technology has major potential to improve the speed and sensitivity of diagnosis and consequent management of patients with this syndrome.