01930nas a2200337   4500000000100000008004100001653001100042653002300053653002400076653001900100653001100119653001200130653002800142653002500170653003000195653003200225653002300257653002700280653000900307653001100316100001300327700001400340700001400354700001300368700001400381245009300395300001100488490000800499520107100507022001401578       2004                            d10aBiopsy10aBacterial Proteins10aAntigens, Bacterial10aDNA, Bacterial10aHumans10aleprosy10aMolecular Sequence Data10aMycobacterium leprae10apolymerase chain reaction10aSensitivity and Specificity10aSequence Alignment10aSequence Analysis, DNA10aSkin10aUganda1 aKramme S1 aBretzel G1 aPanning M1 aKawuma J1 aDrosten C00aDetection and quantification of Mycobacterium leprae in tissue samples by real-time PCR.  a189-930 v1933 a<p>Real-time PCR technology has improved molecular diagnostics of many pathogens, but no such test is available for Mycobacterium leprae. In this report we describe the establishment and the pre-clinical evaluation of such an assay. The test achieved a theoretical analytical sensitivity limit of 194 M. leprae cells per skin biopsy specimen and facilitated quantification of mycobacteria in tissue over a range of 54-54,000,000 cells per sample. In punch skin biopsies from 39 untreated Ugandan patients with newly diagnosed leprosy, the clinical diagnosis could be confirmed in 88.9% of multibacillary and 33.3% of paucibacillary (microscopically negative) patients. Real-time detection thus did not increase the clinical sensitivity of PCR as compared to conventional protocols, in spite of its evidently high analytical sensitivity. On the other hand, as still no culture system exists for M. leprae, the assay appears to be a robust tool for detection of the bacterium in selected clinical situations, as well as for quantitation in experimental settings.</p>
  a0300-8584