{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T13:22:55Z","timestamp":1791638575378,"version":"4.3.4"},"reference-count":37,"publisher":"Hindawi Limited","license":[{"start":{"date-parts":[[2013,1,1]],"date-time":"2013-01-01T00:00:00Z","timestamp":1356998400000},"content-version":"unspecified","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"funder":[{"name":"Research University Cluster"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["BioMed Research International"],"published-print":{"date-parts":[[2013]]},"abstract":"<jats:p>The magnitude of shigellosis in developing countries is largely unknown because an affordable detection method is not available. Current laboratory diagnosis of<jats:italic>Shigella<\/jats:italic>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<jats:italic>invC<\/jats:italic>for<jats:italic>Shigella<\/jats:italic>genus,<jats:italic>rfc<\/jats:italic>for<jats:italic>S. flexneri<\/jats:italic>,<jats:italic>wbgZ<\/jats:italic>for<jats:italic>S. sonnei<\/jats:italic>, and<jats:italic>rfpB<\/jats:italic>for<jats:italic>S. dysenteriae<\/jats:italic>, as well as one internal control (<jats:italic>ompA<\/jats:italic>) gene, was developed in a single reaction to detect and differentiate<jats:italic>Shigella<\/jats:italic>spp. Validation with 120<jats:italic>Shigella<\/jats:italic>strains and 37 non-<jats:italic>Shigella<\/jats:italic>strains yielded 100% specificity. The sensitivity of the PCR was 100\u2009pg of genomic DNA, 5.4 \u00d7 10<jats:sup>4<\/jats:sup>\u2009CFU\/ml, or approximately 120\u2009CFU 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-<jats:italic>Shigella<\/jats:italic>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<jats:italic>Shigella<\/jats:italic>genus and the three<jats:italic>Shigella<\/jats:italic>species responsible for the majority of shigellosis cases.<\/jats:p>","DOI":"10.1155\/2013\/412370","type":"journal-article","created":{"date-parts":[[2013,2,13]],"date-time":"2013-02-13T21:01:49Z","timestamp":1360789309000},"page":"1-9","source":"Crossref","is-referenced-by-count":39,"title":["A Pentaplex PCR Assay for the Detection and Differentiation of<i>Shigella<\/i>Species"],"prefix":"10.1155","volume":"2013","author":[{"given":"Suvash Chandra","family":"Ojha","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology & Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, 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