Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples
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 diffe...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2016-07-01
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| Series: | Brazilian Journal of Infectious Diseases |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1413867016300770 |
| _version_ | 1819024274986369024 |
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| author | Ivano de Filippis Claudia Ferreira de Andrade Nathalia Caldeira Aline Carvalho de Azevedo Antonio Eugenio de Almeida |
| author_facet | Ivano de Filippis Claudia Ferreira de Andrade Nathalia Caldeira Aline Carvalho de Azevedo Antonio Eugenio de Almeida |
| author_sort | Ivano de Filippis |
| collection | DOAJ |
| description | 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. Keywords: Diagnostic, Neisseria meningitidis, Streptococcus pneumoniae, Haemophilus influenzae |
| first_indexed | 2024-12-21T04:52:11Z |
| format | Article |
| id | doaj.art-bfeb223ba64b4f6088b809d0b0dcd1af |
| institution | Directory Open Access Journal |
| issn | 1413-8670 |
| language | English |
| last_indexed | 2024-12-21T04:52:11Z |
| publishDate | 2016-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Brazilian Journal of Infectious Diseases |
| spelling | doaj.art-bfeb223ba64b4f6088b809d0b0dcd1af2022-12-21T19:15:25ZengElsevierBrazilian Journal of Infectious Diseases1413-86702016-07-01204335341S1413-86702016000400335Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samplesIvano de Filippis0Claudia Ferreira de Andrade1Nathalia Caldeira2Aline Carvalho de Azevedo3Antonio Eugenio de Almeida4Corresponding author.; Fundação Oswaldo Cruz (FIOCRUZ), Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz (FIOCRUZ), Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz (FIOCRUZ), Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz (FIOCRUZ), Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz (FIOCRUZ), Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Rio de Janeiro, RJ, BrazilBackground: 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. Keywords: Diagnostic, Neisseria meningitidis, Streptococcus pneumoniae, Haemophilus influenzaehttp://www.sciencedirect.com/science/article/pii/S1413867016300770 |
| spellingShingle | Ivano de Filippis Claudia Ferreira de Andrade Nathalia Caldeira Aline Carvalho de Azevedo Antonio Eugenio de Almeida Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples Brazilian Journal of Infectious Diseases |
| title | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples |
| title_full | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples |
| title_fullStr | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples |
| title_full_unstemmed | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples |
| title_short | Comparison of PCR-based methods for the simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in clinical samples |
| title_sort | comparison of pcr based methods for the simultaneous detection of neisseria meningitidis haemophilus influenzae and streptococcus pneumoniae in clinical samples |
| url | http://www.sciencedirect.com/science/article/pii/S1413867016300770 |
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