Identification and semiquantitation of Mycobacterium avium using a competitive PCR method.
A competitive PCR method with standard DNA (MIMIC) was developed for the rapid detection and semiquantitation of Mycobacterium avium (M. avium) using primers specific for the alpha antigen sequence of the bacteria. DNA from both M. avium and Mycobacterium marinum was amplified by polymerase chain reaction (PCR), but only M. avium could be detected by subsequent blotting confirmation with a probe specific for the bacteria. With the PCR and subsequent dot blot hybridization, as little as 10 fg of the M. avium DNA could be detected, equivalent to about 2 cells of the mycobacteria. In addition, we could distinguish 10(5) CFUs of M. avium from 10(4) CFUs or less by competitive PCR using a MIMIC. The present competitive PCR test enabled rapid identification and semiquantitation of M. avium, and could be used clinically to monitor disease severity and response to treatment of human M. avium disease.
Hashimoto
T
T
Department of Infectious Disease, Kyoto University Hospital, Japan.
Suzuki
K
K
D015373:Bacterial Typing Techniques; D015139:Blotting, Southern; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D006801:Humans; D009161:Mycobacterium / Q000145:classification / Q000235:genetics; D009164:Mycobacterium Infections / Q000382:microbiology; D015269:Mycobacterium avium Complex / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D015270:Mycobacterium avium-intracellulare Infection / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013045:Species Specificity
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial