RESUMO
There are several convenient and accurate molecular assays to detect respiratory bacterial infection. The NeoPlex RB-8 detection kit (NeoPlex RB-8) is a new multiplex real-time PCR assay that simultaneously detects Streptococcus pneumoniae, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, Haemophilus influenzae, Bordetella pertussis, Bordetella parapertussis, and Moraxella catarrhalis in a single test. This study compared the clinical concordance of NeoPlex RB-8 with another method, Seeplex PneumoBacter ACE detection assay (Seeplex PB ACE), which simultaneously detects S. pneumoniae, M. pneumoniae, C. pneumoniae, L. pneumophila, H. influenzae, and B. pertussis We tested 2,137 nasopharyngeal swab and sputum specimens using both assays. For discordant Bordetella parapertussis and M. catarrhalis specimens, we also performed bidirectional sequencing. For S. pneumoniae, M. pneumoniae, C. pneumoniae, L. pneumophila, H. influenzae, and B. pertussis, which are detected by both NeoPlex RB-8 and Seeplex PB ACE, the positive and negative agreement between the two assays ranged from 91.7 to 100% (κ = 0.918 to 1). S. pneumoniae and H. influenzae were the most discordant targets and measured with higher sensitivity and specificity by NeoPlex RB-8 than Seeplex PB ACE. For Bordetella parapertussis and M. catarrhalis, which are not detected by Seeplex PB ACE, NeoPlex RB-8 sensitivity and specificity were >99%. Overall, NeoPlex RB-8 was highly comparable to Seeplex PB ACE, but NeoPlex RB-8 was more clinically accurate, with higher throughput and more convenience.
Assuntos
Bactérias/classificação , Infecções Bacterianas/diagnóstico , Reação em Cadeia da Polimerase Multiplex/normas , Kit de Reagentes para Diagnóstico/normas , Infecções Respiratórias/microbiologia , Bactérias/patogenicidade , Infecções Bacterianas/microbiologia , Humanos , Reação em Cadeia da Polimerase Multiplex/métodos , Nasofaringe/microbiologia , Infecções Respiratórias/diagnóstico , Sensibilidade e Especificidade , Escarro/microbiologiaRESUMO
PURPOSE: Human papillomavirus (HPV) infection in women is known to promote the development of cervical neoplasia. Specific HPV genotypes are more highly associated with disease, and therefore detection and genotyping of HPV infection is critical for preventing and effectively treating cervical cancer. Consequently, various assays using diverse technologies have been developed to detect HPV genotype. Recently the OmniPlex-HPV and GeneFinder HPV methods, based on PCR and Luminex xMAP liquid bead microarray technologies, were developed for the detection of 40 and 32 HPV genotypes, respectively. The purpose of this study was to compare the clinical performance of OmniPlex-HPV and GeneFinder HPV. METHODOLOGY: The study included 300 cytology-confirmed cervical swab specimens. In cases where there was a discrepancy between the two assay results, type-specific direct sequencing was performed. RESULT: We found a high overall agreement between OmniPlex-HPV and GeneFinder HPV for detecting the presence or absence of high-risk HPV (HR HPV) (90.7â%, κ=0.810). However, OmniPlex-HPV showed greater sensitivity than GeneFinder HPV in the identification of multiple genotype-infected samples. Specifically, diagnostic sensitivities for HR HPV positivity in high-grade squamous intra-epithelial lesions (HSIL) were 100.0â% for OmniPlex-HPV and 96.8â% for GeneFinder HPV. CONCLUSIONS: Our results suggest that OmniPlex-HPV and GeneFinder HPV are highly comparable for the detection and genotyping of HPV, but OmniPlex-HPV displays greater accuracy in cases of multiple HPV infection.
Assuntos
Colo do Útero/virologia , Técnicas Citológicas/métodos , Papillomaviridae/isolamento & purificação , Infecções por Papillomavirus/virologia , Reação em Cadeia da Polimerase/métodos , Adulto , Colo do Útero/patologia , DNA Viral/genética , Feminino , Genótipo , Humanos , Papillomaviridae/classificação , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/patologia , Adulto JovemRESUMO
To investigate the underlying genetic influences of primary glaucoma in Korea, molecular analysis was performed in 112 sporadic cases, and results compared with healthy controls. The myocilin (MYOC) and optineurin (OPTN) genes were directly sequenced in 112 unrelated patients, including 17 with primary openangle glaucoma, 19 with juvenile openangle glaucoma, and 76 with normal tension glaucoma. Healthy unrelated Korean individuals (n=100) were used as the nonselected population control. A total of three MYOC and four OPTN variants potentially associated with primary glaucoma were identified in 4 and 18 patients, respectively. A novel variant of MYOC, p.Leu255Pro, was predicted to be potentially pathogenic by in silico analysis. Another, p.Thr353Ile, has been previously reported. These two missense variants were detected in patients with a family history of glaucoma. Combined heterozygous variants p.[Thr123=;Ile288=] were identified in 2 of 112 (2%) patients but not in healthy controls. Among OPTN variants, a novel variant p.Arg271Cys was identified. Homozygous p.[Thr34=;Thr34=] (4/112, 4%), homozygous p.[Met98Lys;Met98Lys] (4/112, 4%), or combined heterozygous p.[Thr34=;Arg545Gln] (9/112, 8%) was significantly associated with the development of primary glaucoma [odds ratio (OR)=8.768, 95% confidence interval (CI)=1.97238.988; relative risk=1.818, 95% CI=1.4732.244; P=0.001]. The present study provides insight into the genetic or haplotype variants of MYOC and OPTN genes contributing to primary glaucoma. Haplotype variants identified in the present study may be regarded as potential contributing factors of primary glaucoma in Korea. Further studies, including those on additional genes, are required to elucidate the underlying pathogenic mechanism using a larger cohort to provide additional statistical power.