Publication: A highly sensitive quantitative real-time PCR assay based on the groEL gene of contemporary Thai strains of Orientia tsutsugamushi
Issued Date
2009-01-01
Resource Type
ISSN
14690691
1198743X
1198743X
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2-s2.0-65949110860
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Mahidol University
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SCOPUS
Bibliographic Citation
Clinical Microbiology and Infection. Vol.15, No.5 (2009), 488-495
Suggested Citation
D. H. Paris, N. Aukkanit, K. Jenjaroen, S. D. Blacksell, N. P.J. Day A highly sensitive quantitative real-time PCR assay based on the groEL gene of contemporary Thai strains of Orientia tsutsugamushi. Clinical Microbiology and Infection. Vol.15, No.5 (2009), 488-495. doi:10.1111/j.1469-0691.2008.02671.x Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/28275
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Title
A highly sensitive quantitative real-time PCR assay based on the groEL gene of contemporary Thai strains of Orientia tsutsugamushi
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Abstract
Partial nucleotide sequences (459bp) of the groEL gene (encoding the 60-kDa heat shock protein, HSP60) from 23 contemporary isolates of Orientia tsutsugamushi isolated from patients with acute scrub typhus in Thailand were compared with 16 reference strain sequences to evaluate the potential of groEL as a conserved and representative target for molecular diagnostics. Overall nucleotide identity within all available O. tsutsugamushi isolates (n = 39) was 98.8% (range: 95.0-100), reflecting a high degree of conservation; nucleotide identities were 67.5% and 65.6%, respectively, when typhus and spotted fever group rickettsiae were included. A highly sensitive and quantitative real-time PCR assay was designed and evaluated using 61 samples, including buffy coats from patients in Thailand and Laos. Reliable and accurate quantitation of bacterial loads allows further investigation of other diagnostic methods and may lead to an improved understanding of the pathophysiology of acute scrub typhus, an important but under-recognized disease. © 2009 The Authors Journal compilation © 2009 European Society of Clinical Microbiology and Infectious Diseases.