五日热巴尔通体RPA检测方法的建立

Real-time Recombinase Polymerase Amplifi cation Assay for Rapid and Specifi c Detection of Bartonella quintana

五日热巴尔通体(B.quintana)病原通过人虱的叮咬在人群中传播,导致间歇性发热,头痛及杆菌样血管瘤等临床症状。本研究以五日热巴尔通体gltA为靶标基因,建立重组聚合酶快速扩增方法(RPA)。通过与Bartonella henselae,Bartonella tribocorum,Bartonella vinsonii,Bartonella washoensis及Bartonella grahamii等不同菌种进行对比检测,证实本研究建立的RPA检1测方法仅特异性识别五日热巴尔通体,无种间无交叉反应,该方法检测敏感性为10 copies/μL,与实时荧光PCR检测敏感性相同。因此,本研究建立的五日热巴尔通体RPA检测方法具备高特异性及敏感性的特点,对五日热巴尔通体菌的检测及疾病临床诊断具有重要意义。

Trench fever characterized by sudden onset with fever and headache is transmitted by a body louse harboring the causative agent Bartonellaquintana.Rapid and sensitive detection of the causative pathogen is essential for timely targeted therapy.In this study,we developed a real-time recombinase polymerase amplification(RPA)assay for the fast detection ofB.quintana.The detection limit was 110 copies/μL,which was the same as real-time RCR.There was no cross-reaction with other Bartonella species(Bartonella henselae,Bartonella tribocorum,Bartonella vinsonii,Bartonella washoensis and Bartonella grahamii).Therefore,it was concluded that the RPA method developed in the present study was efficient in detection of Bartonella quintana infection and could be used for clinical diagnosis of bartonellosis.