Use of polymerase chain reaction to assess efficacy of leprosy chemotherapy.
The assessment of chemotherapy efficacy in leprosy is difficult, since the only reliable method for determining whether the causative organism, Mycobacterium leprae, is viable depends on its growth in mouse foot pads. In an attempt to replace this expensive, time-consuming test, methods based on the polymerase chain reaction (PCR) have been developed. These methods depend on detection of DNA, which is more susceptible to degradation on cell death than are other cell components, so should be a more accurate indicator of viability. We have used a specific PCR assay to detect M leprae DNA in skin biopsy samples from leprosy patients. By use of limiting dilution PCR (LD-PCR), the concentration of M leprae DNA in the original sample could be measured. The DNA concentration was more closely correlated with the morphological index (derived from a staining technique that distinguishes morphologically intact and damaged bacteria) than with the number of bacteria visible (bacterial index, BI, which counts both alive and dead bacteria). In a longitudinal study of multibacillary patients on multi-drug therapy, skin biopsy samples were collected before treatment and 3, 6, 12, and 24 months after the start of therapy. While the BI showed little or no change during treatment, the number of genomes detected by PCR fell sharply, in parallel with the MI. We propose that PCR can be used as a rapid measure of M leprae viability and that this approach can be used for monitoring individual leprosy patients and for assessment of existing and new regimens. The method may be applicable to other infectious diseases in which culture of the causative organism is slow or impossible.
Jamil
S
S
Department of Clinical Sciences, London School of Hygiene and Tropical Medicine, UK.
Keer
J T
JT
Lucas
S B
SB
Dockrell
H M
HM
Chiang
T J
TJ
Hussain
R
R
Stoker
N G
NG
D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D007917:Leprostatic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D007918:Leprosy / Q000188:drug therapy* / Q000382:microbiology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000187:drug effects* / Q000235:genetics; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial; D007917:Leprostatic Agents