alexa Comparison of two different PCR amplification products (the 18-kDa protein gene vs. RLEP repetitive sequence) in the diagnosis of Mycobacterium leprae.
Genetics

Genetics

Molecular Biology: Open Access

Author(s): Kang TJ, Kim SK, Lee SB, Chae GT, Kim JP

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Abstract To determine the best molecular method for diagnosing leprosy, two sets of Mycobacterium leprae-specific primers were compared. Fresh biopsies and slit skin smear samples were obtained from 67 leprosy patients and examined by touchdown (TD) PCR using primers amplifying either a 129-bp fragment of the RLEP repetitive sequence or a 360-bp fragment of the 18-kDa protein gene of M. leprae. Seventeen of 30 (56.7\%) biopsy specimens and four of 37 (10.8\%) slit skin smear specimens were positive using the primer for the 18-kDa protein gene, whereas 24 of 30 (80\%) biopsy and 27 of 37 (73\%) slit skin smear samples showed detectable PCR products in the RLEP repetitive sequence. Twenty-one of 31 cases (67.7\%) with a bacterial index of zero were PCR positive for the primer RLEP repetitive sequence. These results demonstrate that detection of M. leprae using PCR with primers to a RLEP sequence is more sensitive and specific than PCR with the 18-kDa protein gene primers and also slit smears with acid fast staining. PCR of RLEP repetitive sequences is therefore a useful means of detecting M. leprae DNA even when it is present at very low levels.
This article was published in Clin Exp Dermatol and referenced in Molecular Biology: Open Access

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