02334nas a2200409   4500000000100000008004100001260001300042653001500055653001000070653000900080653001900089653001000108653002100118653001600139653001800155653001100173653001100184653001200195653000900207653001600216653002500232653003100257653005200288653002400340653002100364653003200385653000900417100001200426700001100438245012800449856005100577300000900628490000700637050003200644520123400676022001401910       2004                            d  c2004 Mar10aAdolescent10aAdult10aAged10aBiopsy, Needle10aChild10aChild, Preschool10aColorimetry10aCulture Media10aFemale10aHumans10aleprosy10aMale10aMiddle Aged10aMycobacterium leprae10aNucleic Acid Hybridization10aReverse Transcriptase Polymerase Chain Reaction10aRNA, Ribosomal, 16S10aSampling Studies10aSensitivity and Specificity10aSkin1 aHaile Y1 aRyon J00aColorimetric microtitre plate hybridization assay for the detection of Mycobacterium leprae 16S rRNA in clinical specimens.  uhttps://leprosyreview.org/article/75/1/04-0049  a40-90 v75  aInfolep Library - available3 a<p>We have developed a colorimetric microtitre plate hybridization assay in order to simplify detection of Mycobacterium leprae in clinical specimens. This system detects the products amplified by a sensitive RT-PCR assay targeting a species-specific sequence of the bacterial 16S rRNA. The assay detected as few as 10 bacilli isolated from infected nude mouse lymph nodes or human skin biopsies. Sensitivity for diagnosis of clinical specimens was assessed for 58 tissue biopsies from untreated leprosy patients. The assay detected M. leprae RT-PCR products in 100% of biopsies from patients with multibacillary disease and 80% of biopsies from patients with paucibacillary disease, for an overall sensitivity of 91.3%. The test was highly specific as no RT-PCR products were amplified from skin biopsies of normal individuals or patients with skin diseases other than leprosy. The colorimetric assay is faster, more sensitive, and simplifies detection of RT-PCR products compared to Southern blot analysis. It may be useful for diagnosis of difficult cases of leprosy, and, since RNA is rapidly degraded after cell death, it may be appropriate for assessing response to therapy and for distinguishing relapse from reaction.</p>  a0305-7518