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. 2014 Mar;90(3):524-9.
doi: 10.4269/ajtmh.13-0659. Epub 2014 Feb 3.

Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy

Wen Yan  1 , Yan Xing, Lian Chao Yuan, Rong De Yang, Fu Yue Tan, Ying Zhang, Huan-Ying Li
Affiliations

Application of RLEP real-time PCR for detection of M. leprae DNA in paraffin-embedded skin biopsy specimens for diagnosis of paucibacillary leprosy

Wen Yan et al. Am J Trop Med Hyg. 2014 Mar.

Abstract

The TaqMan real-time polymerase chain reaction (PCR) assay was evaluated systematically with respect to the standard curve, linear range, and used for detecting Mycobacterium leprae DNA in paraffin-embedded skin biopsy specimens from 60 confirmed leprosy patients and three healthy individuals and 29 other dermatoses and bacterial DNA from 21 different species. The test was further evaluated with 51 paucibacillary (PB) patients. The results showed that the test had good sensitivity (8 fg) and good specificity with no cross-reactivity with 21 other bacterial species and the control specimens, except one with Xanthomatosis. The real-time PCR detection rate for the 51 PB specimens was 74.5% (38 of 51). We conclude that the real-time PCR test is a useful adjunct test for diagnosing early stage or PB leprosy cases.

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Figures

Figure 1.
Figure 1.
Standardization of the real-time polymerase chain reaction (PCR) assay. Ten to 107 copies of the plasmid DNA were used for standard curve preparation. Ct = −3.44 log + 36.65; R2 = 0.9995.
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