A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species
Joint Authors
Ojha, Suvash Chandra
Yean Yean, Chan
Ismail, Asma
Banga Singh, Kirnpal-Kaur
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-02-13
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
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, and rfpB for S.
dysenteriae, as well 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.
American Psychological Association (APA)
Ojha, Suvash Chandra& Yean Yean, Chan& Ismail, Asma& Banga Singh, Kirnpal-Kaur. 2013. A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species. BioMed Research International،Vol. 2013, no. 2013, pp.1-9.
https://search.emarefa.net/detail/BIM-1004253
Modern Language Association (MLA)
Ojha, Suvash Chandra…[et al.]. A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species. BioMed Research International No. 2013 (2013), pp.1-9.
https://search.emarefa.net/detail/BIM-1004253
American Medical Association (AMA)
Ojha, Suvash Chandra& Yean Yean, Chan& Ismail, Asma& Banga Singh, Kirnpal-Kaur. A Pentaplex PCR Assay for the Detection and Differentiation of Shigella Species. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-9.
https://search.emarefa.net/detail/BIM-1004253
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1004253