摘要
目的建立鉴别诊断恶性疟原虫(P.f)和间日疟原虫(P.v)的多重巢式PCR法。方法针对P.f、P.v 18S rRNA基因设计外引物和内引物,优化引物浓度与退火温度,建立可扩增出两种疟原虫基因片段的多重巢式PCR,并检测54例疑似疟疾临床标本,以镜检法为金标准评价敏感性和特异性等指标。结果该方法可扩增出162 bp(P.f)和112 bp(P.v)基因片段,并能检出混合感染。该方法检测P.f,敏感性为87.50%、特异性为63.33%;检测P.v,敏感性为69.23%、特异性为68.29%。结论所建立的多重巢式PCR方法能可靠诊断疟疾并鉴别虫种,敏感性高,在混合感染的诊断方面具有优越性。
Objective To establish a multiplex-nested PCR for differential detection of Plasmodiumfalciparum （P.f） and Plasmodium vivax （P. v）. Methods The outer and inner primers of PCR were designed according to the sequences of 18S rRNA of P.fand P. v. The reaction system was optimized by screening different concentrations of primer and annealing temperatures. The blood samples from 54 suspected patients with malaria were tested by the optimized multiplex-nested PCR. The sensitivity and specificity of the PCR meth- od were evaluated by using microscopy examination as the gold standard. Results The minimum sizes of amplification products of P. f and P. v were 162 bp and 112 bp respectively. The mixed infection was definitely detectable. The sensitivity arid specificity were 87.50% and 63.33% for P.fdetection, and 69.23% and 68.29% for P. v detection. Conclusion The multiplex-nested PCR devel- oped in this study exhibited high sensitivity for the reliable diagnosis and differentiatial diagnosis of malaria, and showed superiority for the diagnosis of mixed infections of Plasmodium.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2014年第8期565-567,共3页
Chinese Journal of Clinical Laboratory Science
基金
国家质量监督检验检疫总局科技计划(2013IK224)
湖南省科技计划(2012SK3301)
长沙市科技计划(K1205032-31)
中南大学中央高校基本科研业务费专项(2013zzts299)
