02065nas a2200325   4500000000100000008004100001260001300042653001200055653001800067653001500085653001900100653002100119653001100140653001200151653000900163653002800172653002500200653004200225653003000267653002400297100001800321700001500339700001700354245010600371856007800477300001100555490000700566520115200573022001401725       1990                            d  c1990 Sep10aAnimals10aBase Sequence10aDNA Probes10aDNA, Bacterial10aGenes, Bacterial10aHumans10aleprosy10aMice10aMolecular Sequence Data10aMycobacterium leprae10aNucleic Acid Amplification Techniques10apolymerase chain reaction10aSpecies Specificity1 aPlikaytis B B1 aGelber R H1 aShinnick T M00aRapid and sensitive detection of Mycobacterium leprae using a nested-primer gene amplification assay.  uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC268077/pdf/jcm00057-0051.pdf  a1913-70 v283 a<p>By using a set of four nested oligonucleotide primers, a two-step polymerase chain reaction assay for the detection and identification of Mycobacterium leprae that does not require the use of radioactivity labeled hybridization probes was developed. The nested-primer procedure amplified a 347-base-pair product from M. leprae genomic DNA. No amplification products were produced from DNAs of 19 other Mycobacterium species, 19 non-Mycobacterium species, mouse cells, or human cells. Minor amplification products were observed with three additional Mycobacterium species, i.e., "M. lufu", M. simiae, and M. smegmatis. These products were easily distinguished from the M. leprae product by size and restriction enzyme cleavage patterns. The assay could amplify the 347-base-pair product from samples containing as little as 3 fg of M. leprae genomic DNA--the amount of DNA in a single bacillus. The assay also amplified target sequences in crude lysates of M. leprae bacilli isolated from tissue biopsy specimens from infected animals and humans. The entire assay, from sample preparation to data analysis, can be completed in less than 8 h.</p>  a0095-1137