Evaluation of Nested-PCR for early and rapid diagnosis of brucellosis in humans
LIU Zhi-guo1, 2, WANG Miao3, TA Na1, LI Xiao-yan1, YU Rui-ping1, MI Jing-chuan1, LI Dan2, CUI Bu-yun2, LI Zhen-jun2
1. Inner Mongolia Autonomous Region Central for Comprehensive Disease Controled and Prevention, Huhhot 010031, China; 2. National Institute of Infectious Diseases Control and Prevention, Chinese Center for Disease Control and Prevention/ Collaborative Innovation Center for Diagnosis and Treatment of Infectious Disease, Beijing 102206, China; 3. Ulanqab Center for Endemic Disease Control and Prevention, Ulanqab 012000, China
Abstract:The purpose of this study was to explore the feasibility of using Nested-PCR for early and rapid detection of brucellosis in humans. In this study, 19 biovars of the genus Brucella standard reference strain and four vaccine strains (M5, A19, S2 and 104M) had serologic cross-reactivity with O. anthropic, E. coli O∶157 and Y. enterocolitica O∶9. We performed Nested-PCR amplification to evaluate the detection specificity of this method. Subsequently, a double dilution (100~10-7) of Brucella 16M DNA was amplified by Nested-PCR to assess the analytical sensitivity of the assay. Finally, a total of 200 DNA samples from clinical human brucellosis samples were assayed to explore the feasibility of Nested-PCR in clinical diagnosis of human brucellosis. Our results confirmed the presence of a specific amplified band in all Brucella species but not in other strains. The analytical sensitivity of this method was at least 10-6 (equal to 43 fg/μL of B. melitensis 16Μ DNA), which was significantly higher than that of BCSP31-PCR. The positive rates of amplification with the first-round and second-round primers were 4.5% (9/200) and 75.5% (151/200), respectively. High concordance was observed between serum agglutination tests and Nested-PCR. Consequently, we suggest that Nested-PCR may used for early screening of human brucellosis, although further improvement of the method is essential.
刘志国, 王妙, 塔娜, 李晓燕, 尉瑞平, 米景川, 李丹, 崔步云, 李振军. Nested-PCR在人间布鲁氏菌病早期快速诊断中的探讨[J]. 中国人兽共患病学报, 2021, 37(2): 188-192.
LIU Zhi-guo, WANG Miao, TA Na, LI Xiao-yan, YU Rui-ping, MI Jing-chuan, LI Dan, CUI Bu-yun, LI Zhen-jun. Evaluation of Nested-PCR for early and rapid diagnosis of brucellosis in humans. Chinese Journal of Zoonoses, 2021, 37(2): 188-192.
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