A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species

Ojha, Suvash Chandra and Chan, Yean Yean and Ismail, Asma and Banga Singh, Kirnpal Kaur (2013) A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species. BioMed Research International, 2013 (412370). pp. 1-9. ISSN 2314-6133

[img]
Preview
PDF
Download (1677Kb) | Preview

    Abstract

    The magnitude of shigellosis in developing countries is largely unknown because an affordable detection method is not available. Current laboratory diagnosis of Shigella spp. is laborious and time consuming and has low sensitivity. Hence, in the present study, a molecular-based diagnostic assay which amplifies simultaneously four specific genes to identify invC for Shigella genus, rfc for S. flexneri, wbgZ for S. sonnei, andrfpB for S. dysenteriae, aswell as one internal control (ompA) gene, was developed in a single reaction to detect and differentiate Shigella spp. Validation with 120 Shigella strains and 37 non-Shigella strains yielded 100% specificity. The sensitivity of the PCR was 100 pg of genomic DNA, 5.4 × 104 CFU/ml, or approximately 120 CFU per reaction mixture of bacteria. The sensitivity of the pentaplex PCR assay was further improved following preincubation of the stool samples in Gram-negative broth. A preliminary study with 30 diarrhoeal specimens resulted in no cross-reaction with other non-Shigella strains tested. We conclude that the developed pentaplex PCR assay is robust and can provide information about the four target genes that are essential for the identification of the Shigella genus and the three Shigella species responsible for the majority of shigellosis cases.

    Item Type: Article
    Subjects: Q Science > QR Microbiology > QR1-502 Microbiology
    R Medicine > R Medicine (General) > R735-854 Medical education. Medical schools. Research
    Divisions: Institut Penyelidikan Perubatan Molekul (Institute for Research in Molecular Medicine INFORMM) > Article
    Kampus Kesihatan (Health Campus) > Pusat Pengajian Sains Perubatan (School of Medical Sciences) > Article
    Koleksi Naib Canselor (Vice Chancellor Collection) > Profesor Dato' Dr. Asma Ismail > Article
    Zoom Profil Pakar (Expert Profile) > Professor Datuk Dr. Asma Ismail (Office of The Vice-Chancellor)
    Depositing User: Mr Noorazilan Noordin
    Date Deposited: 22 Dec 2017 16:07
    Last Modified: 22 Dec 2017 16:07
    URI: http://eprints.usm.my/id/eprint/38094

    Actions (login required)

    View Item
    Share