Costa J M, Durand R, Deniau M, Rivollet D, Izri M, Houin R, Vidaud M, Bretagne S
Laboratoire de Biologie Moléculaire Marcel Dassault, Hôpital américain de Paris, Neuilly sur Seine, France.
J Clin Microbiol. 1996 Jul;34(7):1831-3. doi: 10.1128/JCM.34.7.1831-1833.1996.
A PCR enzyme-linked immunosorbent assay (ELISA) involving the use of bone marrow aspirates (BMA) and blood samples (BS) for the diagnosis of visceral leishmaniasis (VL) in human immunodeficiency virus-infected patients was developed with primers selected from the sequence of the small-subunit rRNA gene and compared with direct examination and in vitro cultivation. The PCR was optimized for routine diagnosis: processing of samples with lysis of erythrocytes without isolation of leukocytes, enzymatic prevention of contamination, internal control of the reaction, and ELISA testing in a microtitration plate hybridization. Of 79 samples (33 BMA and 46 BS) from 77 patients without VL, all the results were negative. Fifty-three samples (9 BMA and 44 BS) were obtained from 13 patients with VL: 6 samples drawn during anti-Leishmania treatment were negative whatever the technique used, and 47 samples (9 BMA and 38 BS) were positive with at least one technique. The sensitivities were 51% (24 of 47), 81% (38 of 47), and 98% (46 of 47) for direct examination, culture, and PCR, respectively. Thus, PCR ELISA is reliable for diagnosing VL in human immunodeficiency virus-infected patients, and blood sampling should be sufficient for the follow-up.
一种利用骨髓穿刺液（BMA）和血液样本（BS）诊断人类免疫缺陷病毒感染患者内脏利什曼病（VL）的聚合酶链反应酶联免疫吸附测定（ELISA）被开发出来，其引物选自小亚基rRNA基因序列，并与直接检查和体外培养进行了比较。该聚合酶链反应针对常规诊断进行了优化：对样本进行红细胞裂解处理而不分离白细胞，通过酶法防止污染，进行反应的内部对照，并在微量滴定板杂交中进行ELISA检测。来自77例无VL患者的79份样本（33份BMA和46份BS），所有结果均为阴性。从13例VL患者中获取了53份样本（9份BMA和44份BS）：在抗利什曼原虫治疗期间采集的6份样本无论采用何种技术检测均为阴性，47份样本（9份BMA和38份BS）至少有一种技术检测呈阳性。直接检查、培养和聚合酶链反应的灵敏度分别为51%（47份中的24份）、81%（47份中的38份）和98%（47份中的46份）。因此，聚合酶链反应ELISA对于诊断人类免疫缺陷病毒感染患者的VL是可靠的，血液采样对于随访应该足够。
