{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T11:15:32Z","timestamp":1790248532288,"version":"4.1.0"},"reference-count":26,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2005,2,14]],"date-time":"2005-02-14T00:00:00Z","timestamp":1108339200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/2.0\/"},{"start":{"date-parts":[[2005,2,14]],"date-time":"2005-02-14T00:00:00Z","timestamp":1108339200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/2.0\/"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Infect Dis"],"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                        <jats:title>Background<\/jats:title>\n                        <jats:p>In Malaysia, <jats:italic>Shigella<\/jats:italic> spp. was reported to be the third commonest bacterial agent responsible for childhood diarrhoea. Currently, isolation of the bacterium and confirmation of the disease by microbiological and biochemical methods remain as the \"gold standard\". This study aimed to detect the prevalence of four <jats:italic>Shigella<\/jats:italic> virulence genes present concurrently, in randomly selected Malaysian strains via a rapid multiplex PCR (mPCR) assay.<\/jats:p>\n                     <\/jats:sec><jats:sec>\n                        <jats:title>Methods<\/jats:title>\n                        <jats:p>A mPCR assay was designed for the simultaneous detection of chromosomal- and plasmid-encoded virulence genes (<jats:italic>set1A<\/jats:italic>, <jats:italic>set1B<\/jats:italic>, <jats:italic>ial<\/jats:italic> and <jats:italic>ipaH<\/jats:italic>) in <jats:italic>Shigella<\/jats:italic> spp. One hundred and ten Malaysian strains (1997\u20132000) isolated from patients from various government hospitals were used. Reproducibility and sensitivity of the assay were also evaluated. Applicability of the mPCR in clinical settings was tested with spiked faeces following preincubation in brain heart infusion (BHI) broth.<\/jats:p>\n                     <\/jats:sec><jats:sec>\n                        <jats:title>Results<\/jats:title>\n                        <jats:p>The <jats:italic>ipaH<\/jats:italic> sequence was present in all the strains, while each of the <jats:italic>set1A<\/jats:italic>, <jats:italic>set1B<\/jats:italic> and <jats:italic>ial<\/jats:italic> gene was present in 40% of the strains tested. Reproducibility of the mPCR assay was 100% and none of the non-<jats:italic>Shigella<\/jats:italic> pathogens tested in this study were amplified. The mPCR could detect 100 colony-forming units (cfu) of shigellae per reaction mixture in spiked faeces following preincubation.<\/jats:p>\n                     <\/jats:sec><jats:sec>\n                        <jats:title>Conclusions<\/jats:title>\n                        <jats:p>The mPCR system is reproducible, sensitive and is able to identify pathogenic strains of shigellae irrespective of the locality of the virulence genes. It can be easily performed with a high throughput to give a presumptive identification of the causal pathogen.<\/jats:p>\n                     <\/jats:sec>","DOI":"10.1186\/1471-2334-5-8","type":"journal-article","created":{"date-parts":[[2005,2,15]],"date-time":"2005-02-15T07:25:23Z","timestamp":1108452323000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":60,"title":["Detection of virulence genes in Malaysian Shigellaspecies by multiplex PCR assay"],"prefix":"10.1186","volume":"5","author":[{"given":"Kwai Lin","family":"Thong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan Ling Ling","family":"Hoe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"SD","family":"Puthucheary","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rohani","family":"Md Yasin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2005,2,14]]},"reference":[{"key":"156_CR1","first-page":"651","volume":"77","author":"KL Kotloff","year":"1999","unstructured":"Kotloff KL, Winickoff JP, Ivanoff B, Clemens JD, Swerdlow DL, Sansonetti PJ, Adak GK, Levine MM: Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. 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