Data/hora: 11/06/2024 09:55:30

Provedor de dados: | 55
País: | Brazil
Título: | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples
Autores: | de Filippis,Ivano; Andrade,Claudia Ferreira de; Caldeira,Nathalia; Azevedo,Aline Carvalho de; Almeida,Antonio Eugenio de.
Data: | 2016-08-01
Ano: | 2016
Palavras-chave: | Diagnostic; Neisseria meningitidis; Streptococcus pneumoniae; Haemophilus influenzae.
Resumo: | Abstract  Background Several in-house PCR-based assays have been described for the detection of bacterial meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical samples. PCR-based methods targeting different bacterial genes are frequently used by different laboratories worldwide, but no standard method has ever been established. The aim of our study was to compare different in-house and a commercial PCR-based tests for the detection of bacterial pathogens causing meningitis and invasive disease in humans.  Methods A total of 110 isolates and 134 clinical samples (99 cerebrospinal fluid and 35 blood samples) collected from suspected cases of invasive disease were analyzed. Specific sets of primers frequently used for PCR-diagnosis of the three pathogens were used and compared with the results achieved using the multiplex approach described here. Several different gene targets were used for each microorganism, namely ctrA, crgA and nspA for N. meningitidis, ply for S. pneumoniae, P6 and bexA for H. influenzae.  Results All used methods were fast, specific and sensitive, while some of the targets used for the in-house PCR assay detected lower concentrations of genomic DNA than the commercial method. An additional PCR reaction is described for the differentiation of capsulated and non-capsulated H. influenzae strains, the while commercial method only detects capsulated strains.  Conclusions The in-house PCR methods here compared showed to be rapid, sensitive, highly specific, and cheaper than commercial methods. The in-house PCR methods could be easily adopted by public laboratories of developing countries for diagnostic purposes. The best results were achieved using primers targeting the genes nspA, ply, and P6 which were able to detect the lowest DNA concentrations for each specific target.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1413-86702016000400335
Editor: | Brazilian Society of Infectious Diseases
Relação: | 10.1016/j.bjid.2016.04.005
Formato: | text/html
Fonte: | Brazilian Journal of Infectious Diseases  v.20 n.4 2016
Fonte: | info:eu-repo/semantics/openAccess