疟疾检测的进展:唾液携带者的筛选和鉴定

Sean Campos, Mikayla Ybarra, Jonathan Madeti, Rebecca A. Garcia
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摘要

疟原虫是一种寄生虫,可感染红细胞,引起疟疾感染后的流感样症状。传统的诊断方法不包括配子细胞的计数或检测,配子细胞可以在肝脏中储存很长一段时间并再循环。这些携带者可能没有症状,但他们可以将感染传播给他人或蚊子。目前,尚未批准在有症状或无症状的全血样本中检测疟原虫配子细胞的诊断测试。因此,我们开发了实时PCR检测疟疾的活跃状态和携带者状态。第一种是传统的筛选测试,可以检测出导致疟疾感染的五种疟原虫中的任何一种。第二种是伴随试验,用于在无症状且筛查试验结果阴性的患者全血样本中区分和定量恶性疟原虫和间日疟原虫配子细胞。筛选结果显示,唾液中可扩增恶性疟原虫、间日疟原虫、卵形疟原虫、疟疾疟原虫和诺氏疟原虫,总检出限为565拷贝/μL。配子体试验显示,恶性疟原虫与间日疟原虫之间无交叉反应,RNA的检出限为1000拷贝/μL。
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Advances in Malaria Testing: Screening and Identification of Carriers from Saliva
Plasmodium is a parasite that can infect red blood cells and cause flu-like symptoms with malaria infection. Traditional diagnostic methods do not include counting or testing for gametocytes, which can reservoir in the liver for long periods of time and recirculate. These carriers may have no symptoms, but they can transmit infection to others or to mosquitos. Currently, no diagnostic tests have been approved to detect Plasmodium gametocytes in either symptomatic or asymptomatic whole blood samples. Therefore, we developed real-time PCR assays to detect active and carrier states of malaria. The first is a traditional screening test that can detect any of the five Plasmodium species that cause malaria infection. The second is a companion test to differentiate and quantitate Plasmodium falciparum and P. vivax gametocytes in samples of whole blood from patients who may be asymptomatic and present negative results from screening tests. The screening test showed amplification of P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi in saliva with an overall detection limit of 565 copies/μL. The gametocyte test showed no cross-reactivity between P. falciparum and P. vivax with a limit of detection of RNA at 1000 copies/μL.
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