军团菌双重PCR检测方法的建立及应用.pdfVIP

2005 3 15 3 Chinese Journal ofHealth LaboratoryTechnology, M ar 2005; Vol 15 No 3 297 军团菌双重 PCR检测方法的建立及应用 王中民, 杨国芬,谭崇阳,王武芳, 孔 伟 (, 100101) [] : PCR: 2, 5SrRNA M ip 2 , 10 4bp 996 bp: 51 8@ 10 cfu/ ,l 6 104 bp 996 bp, 4 104 bp, 4; 71, 5 2, 2 104 bp, 710% : , [] 双重 PCR; 军团菌; 检测 [] R372 [] A [] 1004- 8685( 2005) 03- 0297- 03 Establishmentand application ofduplex PCR assay fordetectingL eg ion ella W ang Zhong-m in, Yang Guo-f en, Tan Chong-yang, Wang W u-f ang, K ong We i ( Sanitation andAnti- epide icTea ofHeadquarters ofGeneralEquip ent, PLA, Beijing 100101, China) [Abstract] Objective:To establish a duplex PCR assay to detect L eg ionella in environ entalwater1 ethods:Two different sets of oligonucleotide pri ers were si ultaneously used to a plify 104 bp fro 16SrRNA gene and 996 bp fro M ip 2 gene1Results:The sensitivity of this assywas518@ 10 cfu/ l, 104 bp and 996 bp frag ents could be a plified fro 6 reference L eg ionella p neum op h ila strains and 104 bp fro non- L eg ionella p neum op h ila strains, no DNA productwas observed in 4 non- L eg ionella strains 1Of71water sa ples, 104 bp and 996 bp frag entswere observed in 5 sa ples, only 104 bp frag ent in 2 sa - ples, the positive ratewas 710% 1Conclusion:This assay has high sensitivity and specificity for detectingL eg ionella p neum op h ila inwaters 1 [Keywords] Duplex PCR; L eg ionella; Detection ( Legionairescdisease, LD) ( BCYE) BCYE [2]