Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting
Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are leading causes of meningitis and acute invasive infections. PCR-based methods are widely used for the diagnosis and surveillance of bacterial pathogens because of their high sensitivity, specificity and high-throughput c...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Future Science Ltd
2023-02-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0089 |
| _version_ | 1811162432685473792 |
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| author | Ivano de Filippis Aline Carvalho de Azevedo Irene de Oliveira Lima Nicolle F Lima Ramos Claudia Ferreira de Andrade Antonio ECC de Almeida |
| author_facet | Ivano de Filippis Aline Carvalho de Azevedo Irene de Oliveira Lima Nicolle F Lima Ramos Claudia Ferreira de Andrade Antonio ECC de Almeida |
| author_sort | Ivano de Filippis |
| collection | DOAJ |
| description | Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are leading causes of meningitis and acute invasive infections. PCR-based methods are widely used for the diagnosis and surveillance of bacterial pathogens because of their high sensitivity, specificity and high-throughput capabilities compared with conventional laboratory methods. This study evaluated a high-resolution melting qualitative PCR analysis method for the simultaneous detection of these three pathogens. The assay has been optimized to detect three species-specific genes of each organism isolated from clinical samples, enabling accurate identification of the etiological agent. The method proved to be highly sensitive and cheaper than the real-time PCR TaqMan® system because it is probe-free; it could be used for the diagnosis of invasive diseases in public health laboratories of developing countries. |
| first_indexed | 2024-04-10T06:29:30Z |
| format | Article |
| id | doaj.art-5c2905bef0e24eb384cc96b8a13a90c8 |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2024-04-10T06:29:30Z |
| publishDate | 2023-02-01 |
| publisher | Future Science Ltd |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-5c2905bef0e24eb384cc96b8a13a90c82023-03-01T11:21:06ZengFuture Science LtdBioTechniques0736-62051940-98182023-02-0174210110610.2144/btn-2022-0089Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution meltingIvano de Filippis0Aline Carvalho de Azevedo1Irene de Oliveira Lima2Nicolle F Lima Ramos3Claudia Ferreira de Andrade4Antonio ECC de Almeida51Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, Brazil1Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, Brazil1Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, Brazil1Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, Brazil1Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, Brazil1Laboratório de Microrganismos de Referência, Instituto Nacional de Controle de Qualidade em Saúde (INCQS), Fundação Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ, 21040-900, BrazilNeisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae are leading causes of meningitis and acute invasive infections. PCR-based methods are widely used for the diagnosis and surveillance of bacterial pathogens because of their high sensitivity, specificity and high-throughput capabilities compared with conventional laboratory methods. This study evaluated a high-resolution melting qualitative PCR analysis method for the simultaneous detection of these three pathogens. The assay has been optimized to detect three species-specific genes of each organism isolated from clinical samples, enabling accurate identification of the etiological agent. The method proved to be highly sensitive and cheaper than the real-time PCR TaqMan® system because it is probe-free; it could be used for the diagnosis of invasive diseases in public health laboratories of developing countries.https://www.future-science.com/doi/10.2144/btn-2022-0089bacterial meningitisinvasive diseasemultiplex qPCR-HRMrapid diagnosis |
| spellingShingle | Ivano de Filippis Aline Carvalho de Azevedo Irene de Oliveira Lima Nicolle F Lima Ramos Claudia Ferreira de Andrade Antonio ECC de Almeida Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting BioTechniques bacterial meningitis invasive disease multiplex qPCR-HRM rapid diagnosis |
| title | Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting |
| title_full | Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting |
| title_fullStr | Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting |
| title_full_unstemmed | Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting |
| title_short | Accurate, fast and cost-effective simultaneous detection of bacterial meningitis by qualitative PCR with high-resolution melting |
| title_sort | accurate fast and cost effective simultaneous detection of bacterial meningitis by qualitative pcr with high resolution melting |
| topic | bacterial meningitis invasive disease multiplex qPCR-HRM rapid diagnosis |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0089 |
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