02257nas a2200349   4500000000100000008004100001260001300042653002300055653001100078653001900089653002400108653001100132653002300143653001200166653002500178653001300203653003000216653003100246653001900277653003200296653000900328653002100337653002200358100001100380700002200391700001900413245013700432300001000569490000700579520130700586022001401893       2009                            d  c2009 Jun10aBacterial Proteins10aBiopsy10aDNA, Bacterial10aHeat-Shock Proteins10aHumans10aLeprostatic Agents10aleprosy10aMycobacterium leprae10aParaffin10apolymerase chain reaction10aReproducibility of Results10aRNA, Messenger10aSensitivity and Specificity10aSkin10aTissue Embedding10aTreatment Outcome1 aLini N1 aShankernarayan NP1 aDharmalingam K00aQuantitative real-time PCR analysis of Mycobacterium leprae DNA and mRNA in human biopsy material from leprosy and reactional cases.  a753-90 v583 a<p>Mycobacterium leprae, the causative agent of leprosy, is uncultivable in defined media. Development of new diagnostic tools which do not depend on growth of bacteria is needed for the early detection of M. leprae and for monitoring the effectiveness of chemotherapy. We used a real-time PCR-based assay to quantify the copy number of bacterial DNA and hsp18 mRNA from 47 leprosy patients using paraffin-embedded biopsy samples. The assay used was specific, sensitive and reproducible. The applicability of this approach in monitoring the chemotherapy of leprosy was examined. A reduction in DNA and mRNA during chemotherapy was observed and hsp18 mRNA could not be detected in patients who underwent 2 years of multidrug therapy (MDT). However, a considerable amount of M. leprae DNA could be detected even after 2 years of MDT. A significant amount of hsp18 mRNA was found in reactional cases as well. This raises important questions regarding the role of bacterial antigens in leprosy reactions and the rationale of omitting antibiotics in the treatment of reactional cases. Results in this study show that real-time PCR could be a better tool for the careful monitoring of bacillary DNA and mRNA in lesions, which will help to improve diagnosis, disease progression and the treatment regimen.</p>  a0022-2615