Preliminary application of visual LAMP in detecting drug resistant gene blaKPC-2 of carbapenem resistant Klebsiella pneumoniae

doi:10.3969/j.issn.1673-8640.2019.01.012

Abstract

Abstract:

Objective To establish a method for detecting drug resistant gene bla_KPC-2 of carbapenem resistant Klebsiella pneumoniae by visual loop-mediated isothermal amplification （LAMP）. Methods A total of 3 pairs of specific LAMP primers were designed,and the LAMP detection system of 25 μL was established and optimized. The specificity and sensitivity were compared with those of polymerase chain reaction （PCR）. Results A method for detecting drug resistant gene bla_KPC-2 of carbapenem resistant Klebsiella pneumoniae by visual LAMP was established,which was used in the rapid detection of bla_KPC-2 at 63 ℃ for 40 min. The specificity was close to that of PCR,and the sensitivity was higher than that of PCR for 10 times. There were 12 isolates detected with bla_KPC-2 by LAMP among 25 imipenem,meropenem or ertapenem resistant isolates,and there were 5 isolates detected with bla_KPC-2 by LAMP among 31 imipenem,meropenem and ertapenem resistant isolates（P=0.009 9）. Conclusions The established LAMP could be used as a method to detect bla_KPC-2,but it could not replace drug susceptibility test in vitro.

Key words: Loop-mediated isothermal amplification, Klebsiella pneumoniae, Carbapenem resistance, bla_KPC-2

CLC Number:

Q503

ZOU Ping, BAO Junfeng, ZANG Jia, ZHANG Ting, XU Yaohui. Preliminary application of visual LAMP in detecting drug resistant gene bla_KPC-2 of carbapenem resistant Klebsiella pneumoniae[J]. Laboratory Medicine, 2019, 34(1): 56-59.

Figures/Tables 4

References 10

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引物	引物序列（5'~3'）	引物长度（bp）
F3	TGGCTTTTCTGCCACCG	17
B3	TGCGAGCCAGCACAGC	16
FIP	TACACACCGATGGAGCCGCC-CCTCGTCGCGGAACCAT	37
BIP	GGCTCAGGCGCAACTGTAAGT-GCAGCAAGAAAGCCCTTGAA	41
LF	TACACACCGATGGAGCCGCC	20
LB	GGCTCAGGCGCAACTGTAAGT	21

引物	引物序列（5'~3'）	引物长度（bp）
F3	TGGCTTTTCTGCCACCG	17
B3	TGCGAGCCAGCACAGC	16
FIP	TACACACCGATGGAGCCGCC-CCTCGTCGCGGAACCAT	37
BIP	GGCTCAGGCGCAACTGTAAGT-GCAGCAAGAAAGCCCTTGAA	41
LF	TACACACCGATGGAGCCGCC	20
LB	GGCTCAGGCGCAACTGTAAGT	21

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