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 language | English |
|---|---|
| Pages (from-to) | 1035-1039 |
| Number of pages | 5 |
| Journal | Journal of Clinical Microbiology |
| Volume | 32 |
| Issue number | 4 |
| Publication status | Published - 1994 Jan 1 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Microbiology
- Microbiology (medical)
Cite this
Detection of leptospiral DNA by PCR. / Kee, Sun-Ho; Kim, I. S.; Choi, M. S.; Chang, W. H.
In: Journal of Clinical Microbiology, Vol. 32, No. 4, 01.01.1994, p. 1035-1039.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Detection of leptospiral DNA by PCR
AU - Kee, Sun-Ho
AU - Kim, I. S.
AU - Choi, M. S.
AU - Chang, W. H.
PY - 1994/1/1
Y1 - 1994/1/1
N2 - 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.
AB - 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.
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M3 - Article
VL - 32
SP - 1035
EP - 1039
JO - Journal of Clinical Microbiology
JF - Journal of Clinical Microbiology
SN - 0095-1137
IS - 4
ER -