Kirchhoff L V, Votava J R, Ochs D E, Moser D R
Department of Internal Medicine, University of Iowa, Iowa City, USA.
J Clin Microbiol. 1996 May;34(5):1171-5. doi: 10.1128/jcm.34.5.1171-1175.1996.
The diagnosis of acute infection with Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, is generally made by detecting parasites by microscopic examination of fresh blood. Although highly specific, this approach often lacks sensitivity. Several years ago, PCR assays for the detection of T. cruzi were described, but the sensitivities and specificities of these tests have not yet been defined precisely. In the present study, we first compared the sensitivities of PCR methods that differ in sample processing as well as in the target sequences that are amplified. Then, we challenged eight mice with T. cruzi, and on 31 days over a 380-day period, we compared the ability of the PCR method with the highest sensitivity to detect parasites in blood with that of microscopic examination. During the acute phase of the infections, parasites were detected on average 3.9 days earlier by the PCR method than by microscopy. Furthermore, the infected mice were consistently positive by the PCR method during the chronic phase, while parasites were intermittently detected by microscopic examination during that period. Overall, among the 248 comparisons, in 84 the PCR method was positive and no parasites were seen by microscopic examination, whereas the reverse was true in only 1 case, a difference that is highly significant. These findings suggest that this approach should be in patients suspected of having acute Chagas' disease. Moreover, the higher sensitivity of the PCR method observed in both the acute and chronic phases of the T. cruzi infections in the mice that we studied indicates that this approach should be useful in evaluating experimental drugs in T. cruzi-infected laboratory animals.
克氏锥虫是一种导致恰加斯病的原生动物寄生虫,对急性克氏锥虫感染的诊断通常是通过对新鲜血液进行显微镜检查来检测寄生虫。尽管这种方法具有高度特异性,但往往缺乏敏感性。几年前,有人描述了用于检测克氏锥虫的聚合酶链反应(PCR)检测方法,但这些检测方法的敏感性和特异性尚未得到精确界定。在本研究中,我们首先比较了在样本处理以及扩增的靶序列方面存在差异的PCR方法的敏感性。然后,我们用克氏锥虫感染了8只小鼠,并在380天内的31天里,将敏感性最高的PCR方法检测血液中寄生虫的能力与显微镜检查的能力进行了比较。在感染的急性期,PCR方法检测到寄生虫的时间平均比显微镜检查早3.9天。此外,在慢性期,感染的小鼠通过PCR方法始终呈阳性,而在此期间显微镜检查只能间歇性地检测到寄生虫。总体而言,在248次比较中,有84次PCR方法呈阳性但显微镜检查未发现寄生虫,而相反的情况仅出现1次,这种差异具有高度显著性。这些发现表明,对于疑似患有急性恰加斯病的患者应采用这种方法。此外,我们在研究的小鼠克氏锥虫感染的急性期和慢性期均观察到PCR方法具有更高的敏感性,这表明这种方法在评估感染克氏锥虫的实验动物中的实验药物时应会很有用。