03115nas a2200457   4500000000100000008004100001260001300042653002700055653001200082653002300094653000900117653001100126653001800137653002300155653001200178653002900190653000900219653002400228653002500252653004200277653003000319653001700349653002200366100001400388700001300402700001500415700001400430700001500444700001200459700001500471700001600486700001000502700001700512700001500529245014300544856004100687300001100728490000700739520189700746022001402643       2001                            d  c2001 Dec10aAdenosine Triphosphate10aAnimals10aBacterial Proteins10aFoot10aHumans10aImmunotherapy10aLeprostatic Agents10aleprosy10aLuminescent Measurements10aMice10aMice, Inbred BALB C10aMycobacterium leprae10aNucleic Acid Amplification Techniques10apolymerase chain reaction10aTime Factors10aTreatment Outcome1 aGupta U D1 aKatoch K1 aSharma R K1 aSingh H B1 aNatragan M1 aSingh D1 aSharma V D1 aChauhan D S1 aDas R1 aSrivastava K1 aKatoch V M00aAnalysis of quantitative relationship between viability determination in leprosy by MFP, ATP bioluminescence and gene amplification assay.  uhttp://ila.ilsl.br/pdfs/v69n4a04.pdf  a328-340 v693 a<p>Two hundred twenty-one untreated, borderline lepromatous/lepromatous (BL/LL) leprosy patients have been investigated for viability by the mouse foot pad method (MFP), adenosine triphosphate (ATP) and polymerase chain reaction (PCR). The biopsies were collected at the beginning of and 12/24 months after treatment. The patient group was treated with a) immunotherapy (BCG/Mw) + MDT; b) MDT + pyrazinamide; c) control MDT; d) MDT + minocycline 100 mg once a month supervised + ofloxacin 400 mg once a month supervised. Biopsies were divided in three parts for use in the mouse foot pad, molecular and ATP investigations. In untreated and treated patients (at 12 and 24 months), there was a general agreement among all three techniques, and PCR and ATP showed higher positivity as compared to MFP. Further, there was good correlation among the viable biomass estimated by bacillary ATP levels, PCR assay and growth in mouse foot pads. The positivity was observed by MFP as well as PCR assay (18-kDa and 36-kDa) from all of the specimens when the ATP content was more than 3.6 pg/million. When the ATP content was below 3.5 pg/million, the positive takes in MFP decreased but the PCR positivity correlated with ATP bioluminescence up to 0.04 pg/million. When the ATP content was even lower, the uptake in the MFP was possibly a matter of chance, while PCR positivity was observed in 96% of the cases. For specimens with undetectable ATP, positivity was seen in 1% of the cases, showing the inability of ATP bioluminescence method to detect low background due to host ATP. PCR signals in some cases could be due to the higher sensitivity of the method or persistence of DNA after bacterial death in some cases. On the whole, the PCR methods even though targeting DNA have shown good correlations with biomass which confirm their usefulness in monitoring therapeutic responses in leprosy.</p>
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