用于马来布鲁线虫的巢式聚合酶链反应（PCR）检测的灵敏度及基于PCR检测中“游离”DNA的意义。

Sensitivity of the nested-polymerase chain reaction (PCR) assay for Brugia malayi and significance of 'free' DNA in PCR-based assays.

作者信息

Cox-Singh J, Pomrehn A S, Wolfe N D, Rahman H A, Lu H Y, Singh B

机构信息

Faculty of Medicine and Health Sciences, University Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia.

出版信息

Int J Parasitol. 2000 Oct;30(11):1177-9. doi: 10.1016/s0020-7519(00)00104-1.

DOI:10.1016/s0020-7519(00)00104-1

PMID:11027784

Abstract

The blood filtration method was used as the gold standard to determine the detection level of simple blood-spot sampling and nested-polymerase chain reaction (PCR) for Brugia malayi. Of 100 samples, 48 were filtration-positive. Of these, 26 had microfilaria counts that were low enough (<1-29 microfilariae/ml) to accurately assess the limit of detection by nested-PCR. Nested-PCR consistently detected B. malayi DNA in samples with > or = 10 microfilariae/ml. Post-filtration, microfilaria-depleted, blood-spots from microfilaria-positive samples were screened by nested-PCR and B. malayi specific 'free' DNA was detected in 51.7% of these samples. There was no evidence for 'free' DNA in microfilaria-negative individuals from this endemic community.

摘要

血液过滤法被用作金标准，以确定用于马来布鲁线虫的简单血斑采样和巢式聚合酶链反应（PCR）的检测水平。在100份样本中，48份为过滤阳性。其中，26份的微丝蚴计数低至足以（<1 - 29条微丝蚴/毫升）准确评估巢式PCR的检测限。巢式PCR始终能在微丝蚴≥10条/毫升的样本中检测到马来布鲁线虫DNA。过滤后，对微丝蚴阳性样本中微丝蚴耗尽的血斑进行巢式PCR筛查，在51.7%的此类样本中检测到了马来布鲁线虫特异性“游离”DNA。在这个流行社区的微丝蚴阴性个体中没有“游离”DNA的证据。