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首页> 外文期刊>Journal of Clinical Microbiology >PCR enzyme-linked immunosorbent assay for diagnosis of leishmaniasis in human immunodeficiency virus-infected patients.
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PCR enzyme-linked immunosorbent assay for diagnosis of leishmaniasis in human immunodeficiency virus-infected patients.

机译：PCR酶联免疫吸附试验可用于诊断人类免疫缺陷病毒感染患者的利什曼病。

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摘要

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.
机译：使用选自以下序列的引物开发了一种PCR酶联免疫吸附测定（ELISA），其中涉及使用骨髓抽吸物（BMA）和血液样本（BS）诊断人免疫缺陷病毒感染的患者的内脏利什曼病（VL）亚基rRNA基因的鉴定，并与直接检查和体外培养进行比较。对PCR进行了常规诊断的优化：在不分离白细胞的情况下用红细胞裂解处理样品，酶促预防污染，反应的内部控制以及微量滴定板杂交中的ELISA测试。在来自77例没有VL的患者的79个样本（33 BMA和46 BS）中，所有结果均为阴性。从13例VL患者中获得了53个样本（9 BMA和44 BS）：无论采用何种技术，在抗利什曼病治疗期间抽取的6个样本均为阴性，而47个样本（9 BMA和38 BS）为阳性，至少有1个样本为阳性技术。直接检测，培养和PCR的敏感性分别为51％（47个中的24个），81％（47个中的38个）和98％（47个中的46个）。因此，PCR ELISA对于诊断人类免疫缺陷病毒感染的患者的VL是可靠的，并且血液采样应足以进行随访。

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