Rapid determination of Chlamydia trachomatis by loop-mediated isothermal amplification

doi:10.3969/j.issn.1673-8640.2017.09.014

Abstract

Abstract:

Objective To establish a novel loop-mediated isothermal amplification（LAMP） for the simple and rapid determination of Chlamydia trachomatis（CT）.Methods Four primers（2 inner primers and 2 outer primers） for LAMP were designed by targeting the cryptic plasmid gene of CT using PrimerExplorer V4 software. The reaction system and conditions of LAMP were optimized,and the sensitivity and specificity were evaluated. Meanwhile,the results of LAMP were compared with those of real-time fluorescence quantitation polymerase chain reaction（PCR）. Results There was no cross reaction with other common bacteria,and the sensitivity was 100 copies/μL. A total of 90 clinical specimens were determined by LAMP. Compared with real-time fluorescence quantitation PCR,40 of 90 cases were positive,and 50 of 90 cases were negative by LAMP. There was a consistency of 100% between LAMP and real-time fluorescence quantitation PCR.Conclusions LAMP targeting the cryptic plasmid gene of CT is successfully established,and it can be as a novel tool for the rapid determination of CT.

Key words: Loop-mediated isothermal amplification, Nucleic acid determination, Chlamydia trachomatis

CLC Number:

R446.61

WANG Xueliang, LIU Fen, JIANG Lingli, BAO Yun, YANG Yixiao, XIAO Yanqun, WANG Hualiang. Rapid determination of Chlamydia trachomatis by loop-mediated isothermal amplification[J]. Laboratory Medicine, 2017, 32(9): 809-812.

Figures/Tables 5

References 17

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引物	引物序列（5'~3'）	长度（bp）
F3	CCCATGCTTTCAGATTTGC	19
B3	TGGCAGTTACTATAATTTACCATAC	25
FIP	CACAATTAGAAGACGATTCCTTCCTAG- CGGTCAAATAGAGCAAG	44
BIP	TGTTGGGAAAAATAGACATGGATCGG- CGGAGAATTTACTAATTTTTGG	48
CT-F	AAGCTTAATTTAATCGCGAATCAGA	25
CT-R	TCTTTGGATAGCTGCTAATGCATGG	25

引物	引物序列（5'~3'）	长度（bp）
F3	CCCATGCTTTCAGATTTGC	19
B3	TGGCAGTTACTATAATTTACCATAC	25
FIP	CACAATTAGAAGACGATTCCTTCCTAG- CGGTCAAATAGAGCAAG	44
BIP	TGTTGGGAAAAATAGACATGGATCGG- CGGAGAATTTACTAATTTTTGG	48
CT-F	AAGCTTAATTTAATCGCGAATCAGA	25
CT-R	TCTTTGGATAGCTGCTAATGCATGG	25