Ali A, Haque Abdul, Haque Asma, Sarwar Y, Mohsin M, Bashir S, Tariq A
Health Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan.
Epidemiol Infect. 2009 Jan;137(1):102-7. doi: 10.1017/S0950268808000654. Epub 2008 Apr 16.
Classically Salmonella enterica serovar Typhi (S. Typhi) is associated with typhoid, a major health problem in developing countries. However, in recent years S. Paratyphi A and Vi-negative variants of S. Typhi have emerged rapidly. We have developed a nested multiplex PCR targeting five different genes for differential diagnosis of typhoidal pathogens which has been optimized to be directly applicable on clinical blood samples. Of 42 multiplex PCR-positive blood samples, 26, nine, and two were Vi-positive S. Typhi, Vi-negative S. Typhi and S. Paratyphi A, respectively, and five patients were found to have mixed infection. Seventeen patients grew Salmonella from blood culture and the remaining 25 were positive in the Salmonella-specific PCR. Tests with several common pathogens confirmed the specificity of the assay. We conclude that the proposed multiplex PCR is rapid, sensitive and specific for the diagnosis of typhoidal pathogens directly from blood samples.
传统上，肠道沙门氏菌伤寒血清型（伤寒杆菌）与伤寒有关，这是发展中国家的一个主要健康问题。然而，近年来，甲型副伤寒沙门氏菌和伤寒杆菌的Vi阴性变体迅速出现。我们开发了一种针对五个不同基因的巢式多重PCR，用于伤寒病原体的鉴别诊断，该方法已优化为可直接应用于临床血液样本。在42份多重PCR阳性血液样本中，分别有26份、9份和2份为Vi阳性伤寒杆菌、Vi阴性伤寒杆菌和甲型副伤寒沙门氏菌，另有5名患者被发现有混合感染。17名患者血培养中培养出沙门氏菌，其余25名患者沙门氏菌特异性PCR呈阳性。对几种常见病原体的检测证实了该检测方法的特异性。我们得出结论，所提出的多重PCR对于直接从血液样本中诊断伤寒病原体是快速、灵敏和特异的。
