DETECTION OF BABESIA-MICROTI BY POLYMERASE CHAIN-REACTION
DETECTION OF BABESIA-MICROTI BY POLYMERASE CHAIN-REACTION
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DOI：
10.1128/jcm.30.8.2097-2103.1992
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发表时间：
1992-08-01
影响因子：
9.4
通讯作者：
CONRAD, PA
中科院分区：
文献类型：
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作者：
PERSING, DH;MATHIESEN, D;CONRAD, PA
人类巴贝斯虫病是由感染红细胞内类似疟疾的原生动物微小巴贝斯虫引起的，最近在新英格兰的居民中被诊断出越来越频繁。由于寄生虫体积小，以及稀疏的寄生虫病，诊断很困难，这是大多数这种病原体感染的特征。我们通过通用引物扩增真核16S样基因的一部分，产生了B.microti特异的DNA序列信息；随后进行了直接的DNA序列分析。根据该序列信息合成了用于聚合酶链式反应(PCR)的特异性引物。这个基于聚合酶链式反应的系统对检测微小巴贝斯虫表现出强烈的偏见，而不是吉伯逊巴贝斯虫，并且不能扩增脊椎动物的DNA。该体系的分析灵敏度约为3个裂殖子。来自马萨诸塞州楠塔基特岛的12名临床诊断和寄生虫学确诊的巴贝斯虫病患者的血液样本在这一程序的盲法试验中呈PCR阳性。因此，DNA扩增可能为人类巴贝斯虫病的诊断提供常规方法的补充，并可能为监测治疗或加强对这种新出现的病原体的流行病学监测提供一种新的手段。
Human babesiosis, which is caused by infection with the intraerythrocytic malarialike protozoan Babesia microti, has recently been diagnosed with increasing frequency in residents of New England. Diagnosis is difficult because of the small size of the parasite and the sparse parasitemia that is characteristic of most infections with this pathogen. We generated B. microti-specific DNA sequence information by universal primer amplification of a portion of the eukaryotic 16S-like gene; this was followed by direct DNA sequence analysis. Specific primers were synthesized on the basis of this sequence information for use in the polymerase chain reaction (PCR). The PCR-based system demonstrates a strong bias for detection of B. microti as opposed to Babesia gibsoni and does not amplify vertebrate DNA. The analytical sensitivity of the system is approximately three merozoites. Blood specimens from 12 patients with clinically diagnosed and parasitologically confirmed babesiosis from Nantucket Island, Mass., were PCR positive in a blinded test of this procedure. Thus, DNA amplification may provide an adjunct to conventional methods for the diagnosis of human babesiosis and may provide a new means of monitoring therapy or enhancing epidemiological surveillance for this emerging pathogen.