河南省1例输入性内脏利什曼病的实验室诊断

Laboratory diagnosis of an imported case of visceral leishmaniasis in Henan Province

目的分析河南省1例输入性内脏利什曼病病例,探讨内脏利什曼病的实验室诊断方法。方法分析患者的流行病学资料和临床资料,镜检观察骨髓涂片中杜氏利什曼原虫无鞭毛体;rK39试纸条检测血清中利什曼原虫抗体;用两对引物K13A-K13B和LITSR-L5.8S分别扩增利什曼原虫动基体DNA和核糖体DNA内转录间隔区基因片段。结果该患者曾去过内脏利什曼病流行区,有不规则发热、脾肿大、全血细胞减少、白蛋白/球蛋白比例倒置等症状,骨髓涂片查见杜氏利什曼原虫无鞭毛体,rK39试纸条检测阳性,两对引物K13A-K13B和LITSR-L5.8S分别扩增出87 bp和285 bp的片段。两片段序列与杜氏利什曼原虫相应序列的相似性分别为94%和100%。结论结合患者的流行病学资料和临床表现以及实验室检测结果,确诊该病例为内脏利什曼病病例,病原体为杜氏利什曼原虫。

Objective To analyze the epidemiological and clinical data of an imported case of visceral leishmaniasis in Henan Province, and explore the method of laboratory diagnosis of kala-azar. Methods The epidemiological and clinical data of an imported visceral leishmaniasis patient were analyzed. Leishmania donovani bodies in bone marrow smears were observed microscopically. The antibody was detected by using rK39 dipstick test strips. Two pairs of specific primers, K13A-K13B and LITSR-L5.8S, were used to amplify kinetoplast DNA and internal transcribed spacer of rDNA of the parasite, respectively. RRee-sults The patient had been in the epidemic area of visceral leishmaniasis, and had symptoms such as irregular fever, splenomegaly, pancytopenia, and inversed ratio of albumin and globulin. The amastigotes of L. donovani were found in the bone marrow smears, and rK39 test strip was positive, and the PCR products of K13A-K13B and LITSR-L5.8S were 87 bp and 285 bp respectively. The similarities of the two fragment sequences to the corresponding sequences of L. donovani were 94% and 100%, respectively. Conclusion The case is diagnosed as visceral leishmaniasis according to the epidemiological data, clinical manifestations and laboratory test results of the patient, and the pathogen is L. donovani.