A quantitative PCR (TaqMan) assay for pathogenic Leptospira spp

BMC Infectious Diseases

David B McKay²  Leonie J Barnett¹  Meegan L Symonds¹  Michael F Dohnt¹  Greg A Smith³  Ina L Smith³  Lee D Smythe¹ 

[1]WHO/FAO/OIE Collaborating Centre for Reference & Research on Leptospirosis, Centre for Public Health Sciences, Queensland Health Scientific Services, Brisbane, Australia

[2]Sippy Downs Drive, University of the Sunshine Coast, Maroochydore, 4558 Queensland, Australia

[3]Public Health Virology, Centre for Public Health Sciences, Queensland Health Scientific Services, Brisbane, Australia

关键词: diagnosis;    real-time PCR;    TaqMan;    Leptospirosis;   

Others  :  1177156
DOI  :  10.1186/1471-2334-2-13

 received in 2002-04-04, accepted in 2002-07-08,  发布年份 2002

【 摘 要 】

Background

Leptospirosis is an emerging infectious disease. The differential diagnosis of leptospirosis is difficult due to the varied and often "flu like" symptoms which may result in a missed or delayed diagnosis. There are over 230 known serovars in the genus Leptospira. Confirmatory serological diagnosis of leptospirosis is usually made using the microscopic agglutination test (MAT) which relies on the use of live cultures as the source of antigen, often performed using a panel of antigens representative of local serovars. Other techniques, such as the enzyme linked immunosorbent assay (ELISA) and slide agglutination test (SAT), can detect different classes of antibody but may be subject to false positive reactions and require confirmation of these results by the MAT.

Methods

The polymerase chain reaction (PCR) has been used to detect a large number of microorganisms, including those of clinical significance. The sensitivity of PCR often precludes the need for isolation and culture, thus making it ideal for the rapid detection of organisms involved in acute infections. We employed real-time (quantitative) PCR using TaqMan chemistry to detect leptospires in clinical and environmental samples.

Results and Conclusions

The PCR assay can be applied to either blood or urine samples and does not rely on the isolation and culture of the organism. Capability exists for automation and high throughput testing in a clinical laboratory. It is specific for Leptospira and may discriminate pathogenic and non-pathogenic species. The limit of detection is as low as two cells.

【 授权许可】

   
2002 Smythe et al; licensee BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.

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