DNA-PCR and RT-PCR for the 18-kDa gene of Mycobacterium leprae to assess the efficacy of multi-drug therapy for leprosy

Abstract

DNA-PCR and reverse transcription (RT)-PCR for the 18-kDa protein of Mycobacterium leprae were used to examine the efficacy of multi-drug therapy (MDT) in leprosy. MDT was administered for 0-24 months. Fourteen (63.6%) of 22 patients showed positive PCR results after treatment for 12 months and the positive results decreased to 30% after 24 months of MDT. These results did not correlate with the bacterial index (BI) or the IgM antibody titre for the phenolic glycolipid (PGL)-1. One-dimensional densitometric analysis of agarose gels from PCR from the longitudinal study showed a gradual reduction of the 360-bp band after 12-24 months of MDT. RT-PCR for mRNA of the 18-kDa protein successfully tracked bacterial RNA changes in the biopsies and confirmed a decrease in the RNA of M. leprae in patients after MDT for 12 months. Thus, DNA-and RT-PCR for the 18-kDa protein of M. leprae are effective in assessing the efficacy of MDT for leprosy.

Molecular Determination of Mycobacterium leprae Viability by Use of Real-Time PCR

“…The detection of M. leprae RNA has been proposed to be a promising tool for the rapid detection and the measurement of the viability of pathogenic mycobacteria, since the degradation of RNA is relatively rapid upon cell death (3,13). A previous study of M. tuberculosis measured levels of M. tuberculosis 85B (alpha antigen) mRNA, 16S rRNA, and IS6110 DNA using RT-PCR of patients' sputa to ascertain whether they could serve as potential markers of a response to chemotherapy (4).…”

Evaluation of Real-Time and Conventional PCR Targeting Complex 85 Genes for Detection of Mycobacterium leprae DNA in Skin Biopsy Samples from Patients Diagnosed with Leprosy

“…Diagnosis of PNL, for example, can be conclusive after detection through PCR of M. leprae DNA from nerve biopsy specimens (6). Also, reverse transcription-PCR can be used as a measure of M. leprae viability and consequently can assess the efficacy of multidrug therapy (4).…”

Comparison of radiorespirometric budemeyer assay with ATP assay and mouse foot pad test in detecting viable <i>Mycobacterium leprae </i>from clinical samples

“…Presence of viable M. leprae after MB-MDT treatment has been cited in the literature. [17][18][19][20] Poor sensitivity of MFP test as a viability detection method in treated cases is also well recognized. It is known that the multiplication of few viable organisms among mostly dead cells is hampered due to immune response mounted by the normal mice 14 and is the reason why immunocompromised mice are preferred in the detection of persisters.…”