Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal of tuberculosis and respiratory diseases · 1998
[Polymerase chain reaction associated with dig-labelled DNA hybridization of mip gene to identify new isolated Legionella pneumophila strains].
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Lu X, Peng X, Ren H
Аннотация
OBJECTIVE: By testing mip gene, one highly conservative virulence gene of Legionella pneumophila (Lp), as targets of polymerase chain reaction (PCR) and Digoxin-labelled probe hybridization to establish a rapid, specific and sensitive gene analysis method that can identify new isolated suspected bacteria strains as Lp.
METHOD: Abstracting all the CDC reference Lp strains' and none-Lp Legionella strains' DNA, as well as the domestic isolated Lp strains' and some non-Legionella control bacteria's DNA as templates, then were processed. All the positive PCR products(if no PCR amplicons were available, the original whole DNA should be detected) were hybridized with Digoxingen-labelled mip gene probe in dot-blot procedure.
RESULT: All the tested Lp strains had positive PCR and hybridization results, at the same time, all none-Legionella bacteria and none-Lp Legionella strains got negative results. 6 of 26 isolates were identified as Lp strains by this method.
CONCLUSION: Such a Lp strain identification procedure shows high specificity and sensitivity (nearly 100%, in this study), and mean-while can be completed in a relative short time. Surely, this method can be largely available and has a potential value in diagnosis of clinical Legionellosis cases and pursuing the pathogen during Legionnaires' disease outbreaks.
METHOD: Abstracting all the CDC reference Lp strains' and none-Lp Legionella strains' DNA, as well as the domestic isolated Lp strains' and some non-Legionella control bacteria's DNA as templates, then were processed. All the positive PCR products(if no PCR amplicons were available, the original whole DNA should be detected) were hybridized with Digoxingen-labelled mip gene probe in dot-blot procedure.
RESULT: All the tested Lp strains had positive PCR and hybridization results, at the same time, all none-Legionella bacteria and none-Lp Legionella strains got negative results. 6 of 26 isolates were identified as Lp strains by this method.
CONCLUSION: Such a Lp strain identification procedure shows high specificity and sensitivity (nearly 100%, in this study), and mean-while can be completed in a relative short time. Surely, this method can be largely available and has a potential value in diagnosis of clinical Legionellosis cases and pursuing the pathogen during Legionnaires' disease outbreaks.