DETECTION OF LEPTOSPIRAL DNA BY PCR

被引:37
作者
KEE, SH [1 ]
KIM, IS [1 ]
CHOI, MS [1 ]
CHANG, WH [1 ]
机构
[1] SEOUL NATL UNIV,COLL MED,DEPT MICROBIOL,28 YONGON DONG,CHONGNO GU,SEOUL 110799,SOUTH KOREA
关键词
D O I
10.1128/JCM.32.4.1035-1039.1994
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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 mul 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.
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页码:1035 / 1039
页数:5
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