Gomes Aparecida H S, Armelin Izabel M, Menon Sueli Z, Pereira-Chioccola Vera L
Laboratorio I - do Instituto Adolfo Lutz, Rua: Julio Hanser, 49, Sorocaba, SP, Brazil.
Exp Parasitol. 2008 Jul;119(3):319-24. doi: 10.1016/j.exppara.2008.02.014. Epub 2008 Mar 14.
Cutaneous leishmaniases present similar clinical appearances, but differing prognosis in the course of infection. Ulcers caused by parasites of the subgenus Viannia are more aggressive than ulcers caused by parasites of the subgenus Leishmania. Another problem is distinguishing between true Leishmania infection and other skin diseases in endemic areas, where cutaneous lesions and a single positive Montenegro intradermal test are enough to submit patients to specific treatment for cutaneous leishmaniasis. This study evaluated the efficacy of PCR in detecting in Leishmania in patients with cutaneous lesions. Leishmania (V.) braziliensis complex was determined by a primer pair from the multicopy spliced leader RNA. The results were compared to those of traditional methods. We analyzed biopsies of 109 patients with cutaneous lesions in the second most endemic region of Sao Paulo State, Brazil. Definitive diagnosis was established by clinical and "consensus laboratory criteria" (positive culture, stained tissue smears or PCR). Of 52 patients with cutaneous leishmaniasis, 96% had positive PCR, 69%, positive parasitological tests and 100%, positive Montenegro intradermal tests. Histopathological examination (only in 32 samples) were positive in 14 samples, suggestive in 14 and negative in 4 samples. All 57 patients with other etiologies had negative results in parasitological methods, PCR and histopathological examination (in 39 samples), but Montenegro intradermal tests were positive in 35%. PCR was highly sensitive and specific for L. (V.) braziliensis complex detection compared with other laboratory methods. Despite the specificity of the parasitological tests, the sensitivity was less than 70%. Montenegro intradermal reaction was highly sensitive, but with low specificity, only 65%. As suggestive results in histopathological examinations were shown in 14 samples, it was difficult to determine the true result. PCR applied to biopsies proved to be useful for differential diagnosis of cutaneous lesions of other etiologies in patients living in endemic areas. The advantages are most striking in clinical specimens with scarce amastigotes for which conventional methods have low sensitivity and should be considered for clinical and epidemiological patterns. On the other hand, both Montenegro intradermal test and parasitological methods are only modestly effective in cutaneous leishmaniasis diagnosis.
皮肤利什曼病临床表现相似,但感染过程中的预后不同。维安亚亚属寄生虫引起的溃疡比利什曼原虫亚属寄生虫引起的溃疡更具侵袭性。另一个问题是在流行地区区分真正的利什曼原虫感染和其他皮肤病,在这些地区,皮肤病变和单次阳性的蒙氏皮内试验足以让患者接受皮肤利什曼病的特异性治疗。本研究评估了聚合酶链反应(PCR)在检测皮肤病变患者利什曼原虫方面的有效性。通过来自多拷贝剪接前导RNA的一对引物确定巴西利什曼原虫(维安亚亚属)复合体。将结果与传统方法的结果进行比较。我们分析了巴西圣保罗州第二高流行地区109例皮肤病变患者的活检样本。通过临床和“共识实验室标准”(阳性培养、染色组织涂片或PCR)确立明确诊断。在52例皮肤利什曼病患者中,96%的患者PCR呈阳性,69%的患者寄生虫学检测呈阳性,100%的患者蒙氏皮内试验呈阳性。组织病理学检查(仅32份样本)中,14份样本呈阳性,14份提示阳性,4份样本呈阴性。所有57例其他病因患者的寄生虫学方法、PCR和组织病理学检查(39份样本)结果均为阴性,但35%的患者蒙氏皮内试验呈阳性。与其他实验室方法相比,PCR对巴西利什曼原虫(维安亚亚属)复合体检测具有高度敏感性和特异性。尽管寄生虫学检测具有特异性,但其敏感性低于70%。蒙氏皮内反应高度敏感,但特异性较低,仅为65%。由于14份样本的组织病理学检查结果提示阳性,因此难以确定真实结果。应用于活检样本的PCR被证明有助于对流行地区其他病因的皮肤病变进行鉴别诊断。在无鞭毛体稀少的临床样本中,其优势最为明显,因为传统方法对这类样本敏感性较低,在临床和流行病学模式中应予以考虑。另一方面,蒙氏皮内试验和寄生虫学方法在皮肤利什曼病诊断中效果都不太理想。