Molecular characterization of serogrouping and virulence genes of Malaysian Vibrio cholerae isolated from different sources.
A pair of primers targeting the hlyA gene for Vibrio cholerae which could distinguish the classical from El Tor biotypes was designed and combined with other specific primers for ompW, rfb complex, and virulence genes such as ctxA, toxR, and tcpI in a multiplex PCR (m-PCR) assay. This m-PCR correctly identified 39 V. cholerae from clinical, water and seafood samples. The efficiency of this multiplex PCR (m-PCR) was compared with conventional biochemical and serogrouping methods. One O139 and 25 O1 V. cholerae strains including 10 environmental strains harbored all virulence-associated genes except 1 clinical strain which only had toxR and hlyA genes. Thirteen environmental strains were classified as non-O1/non-O139 and had the toxR and hlyA genes only. The detection limit of m-PCR was 7 x 10(4) cfu/ml. The m-PCR test was reliable and rapid and reduced the identification time to 4 h.
Shuan Ju Teh
Cindy
C
Institute of Biological Sciences, University of Malaya, Kuala Lumpur, Malaysia.
Lin Thong
Kwai
K
Tein Ngoi
Soo
S
Ahmad
Norazah
N
Balakrish Nair
Gopinath
G
Ramamurthy
Thandavarayan
T
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000379:methods; D002771:Cholera / Q000821:virology*; D017931:DNA Primers; D006460:Hemolysin Proteins / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008296:Malaysia; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017747:Seafood / Q000382:microbiology*; D012623:Seawater / Q000382:microbiology*; D012680:Sensitivity and Specificity; D012703:Serotyping; D014734:Vibrio cholerae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity; D014774:Virulence / Q000235:genetics
D001426:Bacterial Proteins; D017931:DNA Primers; D006460:Hemolysin Proteins; C084287:hlyA protein, Vibrio cholerae