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Development of a melting-curve based multiplex real-time PCR assay for simultaneous detection of Streptococcus agalactiae and genes encoding resistance to macrolides and lincosamides.
Otaguiri, Eliane Saori; Morguette, Ana Elisa Belotto; Morey, Alexandre Tadachi; Tavares, Eliandro Reis; Kerbauy, Gilselena; de Almeida Torres, Rosângela S L; Chaves Júnior, Mauricio; Tognim, Maria Cristina Bronharo; Góes, Viviane Monteiro; Krieger, Marco Aurélio; Perugini, Marcia Regina Eches; Yamauchi, Lucy Megumi; Yamada-Ogatta, Sueli Fumie.
Afiliação
  • Otaguiri ES; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Morguette AEB; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Morey AT; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Tavares ER; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Kerbauy G; Departamento de Enfermagem, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • de Almeida Torres RSL; Laboratory of Bacteriology, Epidemiology Laboratory and Disease Control Division, Laboratório Central do Estado do Paraná - LACEN, Curitiba, PR, Brazil.
  • Chaves Júnior M; Departamento de Medicina, Hospital Universitário de Maringá, Universidade Estadual de Maringá, Maringá, Brazil.
  • Tognim MCB; Departamento de Ciências Básicas da Saúde, Universidade Estadual de Maringá, Maringá, Brazil.
  • Góes VM; Instituto Carlos Chagas, Fundação Instituto Oswaldo Cruz, Curitiba, Brazil.
  • Krieger MA; Instituto Carlos Chagas, Fundação Instituto Oswaldo Cruz, Curitiba, Brazil.
  • Perugini MRE; Departamento de Patologia, Análises Clínicas e Toxicológicas, Centro de Ciências da Saúde, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Yamauchi LM; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil.
  • Yamada-Ogatta SF; Departamento de Microbiologia, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, Paraná, Brazil. ogatta@uel.br.
BMC Pregnancy Childbirth ; 18(1): 126, 2018 May 03.
Article em En | MEDLINE | ID: mdl-29724169
BACKGROUND: Streptococcus agalactiae or Group B Streptococcus (GBS) remains the leading cause of infections in newborns worldwilde. Prenatal GBS screening of pregnant women for vaginal-rectal colonization is recommended in many countries to manage appropriate intrapartum antimicrobial prophylaxis for those identified as carriers. In this study, a novel melting-curve based multiplex real-time PCR assay for the simultaneous detection of GBS and macrolide and lincosamide resistance markers was developed. The usefulness of the assay was evaluated for rapid and accurate prenatal GBS screening. METHODS: One hundred two pregnant women who were at 35-37 weeks of gestation were enrolled in this study. The analytical performance of the multiplex real-time PCR was first tested using a panel of reference and clinical bacterial and fungal strains. To test the clinical performance, vaginal-rectal swabs were obtained from pregnant women who were seen at the teaching hospital for regular prenatal care. The results of real-time were compared with those obtained from microbiological analyses. RESULTS: The real-time PCR assay showed 100% specificity and a limit of detection of 104 colony forming units equivalent per reaction. The prevalence of GBS colonization among the population studied was 15.7% (16/102) based on a positive culture and the real-time PCR results. Agreement between the two assays was found for 11 (68.75%) GBS colonized women. Using the culture-based results as a reference, the multiplex real-time PCR had a sensitivity of 91.7% (11/12, CI 59.7-99.6%), a specificity of 95.5% (86/90, CI 89.8-98.7%), a positive predictive value of 73.3% (11/15, CI 44.8-91.1%) and a negative predictive value of 98.9% (86/87, CI 92.9-99.9%). CONCLUSION: The multiplex real-time PCR is a rapid, affordable and sensitive assay for direct detection of GBS in vaginal-rectal swabs.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diagnóstico Pré-Natal / Streptococcus agalactiae / Portador Sadio / Farmacorresistência Bacteriana Tipo de estudo: Diagnostic_studies / Prognostic_studies / Risk_factors_studies Limite: Female / Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Diagnóstico Pré-Natal / Streptococcus agalactiae / Portador Sadio / Farmacorresistência Bacteriana Tipo de estudo: Diagnostic_studies / Prognostic_studies / Risk_factors_studies Limite: Female / Humans Idioma: En Ano de publicação: 2018 Tipo de documento: Article