Dupouy-Camet J, de Souza S L, Maslo C, Paugam A, Saimot A G, Benarous R, Tourte-Schaefer C, Derouin F
Laboratoire de Parasitologie, Hôpital Cochin, Paris, France.
J Clin Microbiol. 1993 Jul;31(7):1866-9. doi: 10.1128/jcm.31.7.1866-1869.1993.
Detection of Toxoplasma gondii in blood by means of the polymerase chain reaction (PCR) may facilitate the diagnosis and follow-up of cerebral toxoplasmosis in patients with AIDS. We evaluated this approach with seven patients with tissue culture-proven parasitemia, 14 patients with presumptive cerebral toxoplasmosis, and 17 healthy human immunodeficiency virus-positive controls. Each sample of blood was assayed on three different occasions by a PCR assay based on detection of the gene encoding the P30 surface protein. A positive PCR diagnosis required positivity in at least two of the three PCR tests. None of the controls had a positive PCR diagnosis, but six of the seven patients with parasitemia did. Cerebral toxoplasmosis was confirmed in 13 of the 14 patients with a presumptive diagnosis; diagnosis by PCR was positive before treatment for 9 of these 13 patients, whereas tissue culture was positive for only 1 patient. During treatment, blood samples were taken from 14 patients at regular intervals until day 12. PCR diagnosis became negative on ethidium-stained gels, but persistent signals were observed after hybridization, in some cases, for up to 12 days after initiation of therapy. PCR on venous blood could thus be a sensitive and noninvasive method for the diagnosis of cerebral and disseminated toxoplasmosis in AIDS patients and could be a potential tool for monitoring the effects of treatment.
通过聚合酶链反应(PCR)检测血液中的弓形虫可能有助于艾滋病患者脑弓形虫病的诊断及随访。我们对7例经组织培养证实有寄生虫血症的患者、14例疑似脑弓形虫病的患者以及17例健康的人类免疫缺陷病毒阳性对照者采用了这种方法。每份血液样本均基于对编码P30表面蛋白的基因的检测,通过PCR检测在三个不同时间点进行测定。PCR阳性诊断要求在三次PCR检测中至少有两次呈阳性。所有对照者的PCR诊断均为阴性,但7例有寄生虫血症的患者中有6例呈阳性。14例疑似诊断患者中有13例确诊为脑弓形虫病;这13例患者中有9例在治疗前PCR诊断呈阳性,而组织培养仅1例呈阳性。治疗期间,对14例患者定期采集血样直至第12天。在溴化乙锭染色的凝胶上PCR诊断变为阴性,但在杂交后,有些情况下在开始治疗后长达12天仍观察到持续信号。因此,静脉血PCR可能是诊断艾滋病患者脑和播散性弓形虫病的一种敏感且非侵入性的方法,并且可能是监测治疗效果的一种潜在工具。