[Detection and identification method of three mycobacterium species and genus specific detection by polymerase chain reaction and DNA hybridization with alkaline phosphatase labeled oligonucleotide probe].
Rapid detection and identification method of mycobacteria from specimen with infected mycobacteriosis was established by the combination of polymerase chain reaction and alkaline phosphatase labeled oligonucleotide probe (PCR-ALPDH). We prepaired four kinds of probes: Mycobacterium tuberculosis complex, M. avium, M. intracellulare, and mycobacterial-probe other than tuberculosis. PCR-ALPDH was compared with conventional methods using 234 specimens which suspected mycobacteriosis. In culture, 68 specimens (29.1%) were positive and 166 specimens (70.9%) were negative. In PCR-ALPDH, 88 specimens (37.6%) were positive and 146 specimens (62.4%) were negative. In 68 specimens which were positive in culture, the agreement of results of conventional identification and PCR-ALPDH for each probes were: 39/40 (PCR-ALPDH/Culture, 97.5%) in M. tuberculosis 5/9 (55.6%) in M. avium, 6/6 (100%) in M. intracellulare and 22/28 (88.6%) in MOTT isolation. Among 166 negative culture specimens, 27 specimens were positive in PCR-ALPDH. The results indicated that PCR-ALPDH method was applicable for the establishment of rapid and sensitive mycobacterial diagnostic system.
Komatsu
M
M
Department of Clinical Pathology, Tenri Hospital.
Shimakawa
K
K
Aihara
M
M
Matsuo
S
S
Ezaki
T
T
D000469:Alkaline Phosphatase; D001483:Base Sequence; D004269:DNA, Bacterial / Q000032:analysis*; D005260:Female; D006801:Humans; D008875:Middle Aged; D008969:Molecular Sequence Data; D009161:Mycobacterium / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D000469:Alkaline Phosphatase