Detection of leptospiral DNA by PCR

Sun-Ho Kee, I. S. Kim, M. S. Choi, W. H. Chang

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Abstract

An EcoRI fragment (1.2 kb) which is highly conserved among Leptospira interrogans isolated in Korea was cloned into pBluescript vector from L. interrogans serovar lai WH20. The EcoRI fragment was sequenced, and a pair of primers (LP1 and LP2) was designed for PCR assay. PCR amplification of target DNA obtained from cultured L. interrogans showed that 274 bp could be detected when as little as 100 fg of leptospiral genomic DNA was used in the reaction mixture. No amplification of DNA was detected from DNA of Leptospira biflexa serovars patoc and sau paulo, Borrelia burgdorferi, Staphylococcus aureus, Escherichia coli, and Salmonella typhimurium. Amplification of 274- bp target DNA could be detected in DNA samples purified from 500 μl of blood collected from experimentally infected gerbils 2 days after infection, while antibodies to L. interrogans could be detected by the microscopic agglutination test 7 days after infection. The specificity and high sensitivity of the test provided valuable tools for the early diagnosis of leptospirosis.

Original languageEnglish
Pages (from-to)1035-1039
Number of pages5
JournalJournal of Clinical Microbiology
Volume32
Issue number4
Publication statusPublished - 1994 Jan 1
Externally publishedYes

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ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Kee, S-H., Kim, I. S., Choi, M. S., & Chang, W. H. (1994). Detection of leptospiral DNA by PCR. Journal of Clinical Microbiology, 32(4), 1035-1039.