Persing D H, Mathiesen D, Marshall W F, Telford S R, Spielman A, Thomford J W, Conrad P A
Department of Laboratory Medicine and Pathology, Mayo Foundation, Rochester, Minnesota 55905.
J Clin Microbiol. 1992 Aug;30(8):2097-103. doi: 10.1128/jcm.30.8.2097-2103.1992.
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.
人类巴贝斯虫病由红细胞内类似疟原虫的微小巴贝斯虫感染所致,最近在新英格兰地区居民中的诊断频率日益增加。由于该寄生虫体积小且大多数感染病例的寄生虫血症稀少,诊断较为困难。我们通过对真核生物16S样基因的一部分进行通用引物扩增来生成微小巴贝斯虫特异性DNA序列信息,随后进行直接DNA序列分析。基于该序列信息合成了特异性引物,用于聚合酶链反应(PCR)。与吉氏巴贝斯虫相比,基于PCR的系统对微小巴贝斯虫的检测具有很强的偏向性,且不扩增脊椎动物DNA。该系统的分析灵敏度约为三个裂殖子。在对来自马萨诸塞州楠塔基特岛的12例临床诊断且经寄生虫学确诊的巴贝斯虫病患者的血液标本进行的盲法检测中,该检测程序的PCR结果呈阳性。因此,DNA扩增可为人类巴贝斯虫病的传统诊断方法提供辅助手段,并可能为监测治疗或加强对这种新出现病原体的流行病学监测提供新方法。