- JOHN LIBBEY EUROTEXT LTD
We have developed a practical method of detecting Mycobacterium leprae DNA in leprosy patients using the polymerase chain reaction (PCR). We have designed a new set of primer oligonucleotides based on the DNA sequence encoding the M. leprae heat shock protein (hsp) 70. In order to avoid non-specific amplification of human or other bacterial DNA, regions of hsp 70 with low amino acid sequence homologies were selected for PCR primers, which also showed low homologies at the nucleic acid level. Instead of skin biopsy specimens, tissue fluid, which may contain M. leprae, was obtained from a needle inserted in the skin. The specificity of the amplified DNA fragment was confirmed by a simplified detection method, which produced the same results as Southern blot analysis. This study demonstrated useful, practical improvements of the PCR method that enable leprosy patients to be detected in the field.
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