河南省1例输入性皮肤利什曼病的诊断与分析
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摘要
患者,男,2017年6月到乌兹别克斯坦务工,7月2日发现左胸臂部有2个小红疹,回国后于10月15日前后发现红疹变大,有少量脓液。用利什曼病快速检测试纸条检测患者血清利什曼原虫抗体结果为阴性。取皮损处组织镜检未查见利什曼原虫无鞭毛体。取皮损处组织置NNN培养基培养,培养至第8天镜检查见大量前鞭毛体。提取前鞭毛体DNA,用利什曼原虫属特异性引物K13A/K13B和L5.8S/LITSR分别扩增出120和350 bp的片段,片段序列与硕大利什曼原虫相应序列(Gen Bank登录号:EU370906.1和FN677342.1)的同源性分别为90%和98%。确诊患者为输入性皮肤利什曼病,病原体为硕大利什曼原虫。患者经过葡萄糖酸锑钠注射液治疗2个疗程(总剂量7.2 g,每个疗程6 d,两个疗程间隔1周),病情得到控制,现已康复。
The male patient who went to Uzbekistan in June 2017 and found two small red rashes on the left thoracic arm on July 2 returned to China in October. The rashes became larger with a small amount of pus. A rapid strip test for leishmaniasis showed that the patient serum was negative for anti-Leishmania antibody. Microscopic examination of the lesioned tissue revealed no Leishmania amastigotes. The lesioned tissue was cultured in NNN medium and a large number of promastigotes were found in culture on day 8. The promastigote DNA was extracted and underwent PCR amplification using Leishmania-specific primers K13A/K13B and L5.8 S/LITSR, producing fragments of 120 and 350 bp. The two products were 90% and 98% homologous to the corresponding sequences of L.major(Gen Bank accession number: EU370906.1 and FN677342.1). The patient was diagnosed as imported cutaneous leishmaniasis and the pathogen was identified to be L. major. The disease was controlled after 2 treatment courses with sodium stibogluconate at a total dosage of 7.2 g( 6 days for each course, with a 1-week interval between them). The patient had recovered by the end of the study.
The male patient who went to Uzbekistan in June 2017 and found two small red rashes on the left thoracic arm on July 2 returned to China in October. The rashes became larger with a small amount of pus. A rapid strip test for leishmaniasis showed that the patient serum was negative for anti-Leishmania antibody. Microscopic examination of the lesioned tissue revealed no Leishmania amastigotes. The lesioned tissue was cultured in NNN medium and a large number of promastigotes were found in culture on day 8. The promastigote DNA was extracted and underwent PCR amplification using Leishmania-specific primers K13A/K13B and L5.8 S/LITSR, producing fragments of 120 and 350 bp. The two products were 90% and 98% homologous to the corresponding sequences of L.major(Gen Bank accession number: EU370906.1 and FN677342.1). The patient was diagnosed as imported cutaneous leishmaniasis and the pathogen was identified to be L. major. The disease was controlled after 2 treatment courses with sodium stibogluconate at a total dosage of 7.2 g( 6 days for each course, with a 1-week interval between them). The patient had recovered by the end of the study.
引文
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[16]Doudi M,Hejazi SH,Razavi MR,et al.Comparative molecular epidemiology of Leishmania major and Leishmania tropica by PCR-RFLP technique in hyper endemic cities of Isfahan and Bam,Iran[J].Med Sci Monit,2010,16(11):CR530-535.
[17]Bensoussan E,Nasereddin A,Jonas F,et al.Comparison of PCR assays for diagnosis of cutaneous leishmaniasis[J].J Clin Microbiol,2006,44(4):1435-1439.
[18]熊光华,金长发.白蛉栖性及其治理对策的研究进展[J].中国寄生虫学与寄生虫病杂志,2006,24(4):293-298.
[2]Akkafa F,Dilmec F,Alpua Z.Identification of Leishmania parasites in clinical samples obtained from cutaneous leishmaniasis patients using PCR-RFLP technique in endemic region,Sanliurfa province,in Turkey[J].Parasitol Res,2008,103(3):583-586.
[3]Desjeux P.Leishmaniasis:current situation and new perspectives[J].Comp Immunol Microbiol Infect Dis,2004,27(5):305-318.
[4]Amro A,Gashout A,Al-Dwibe H,et al.First molecular epidemiological study of cutaneous leishmaniasis in Libya[J].PLo S Negl Trop Dis,2012,6(6):e1700.
[5]管立人,柴君杰.中国利什曼病的现状和对开展防治工作的若干建议[J].地方病通报,2000,15(3):49-53.
[6]Lun ZR,Wu MS,Chen YF,et al.Visceral leishmaniasis in China:an endemic disease under control[J].Clin Microbiol Rev,2015,28(4):987-1004.
[7]杨玥涛,张敏,高春花,等.两例输入性皮肤利什曼病的诊断与病原体鉴定[J].中国寄生虫学与寄生虫病杂志,2011,29(6):461-464.
[8]赖德华,吴娜,谢祎婷,等.一例输入性皮肤利什曼病病原体的鉴定[J].中国寄生虫学与寄生虫病杂志,2016,34(3):295-297.
[9]Waki K,Dutta S,Ray D,et al.Transmembrane molecules for phylogenetic analyses of pathogenic protists:Leishmaniaspecific informative sites in hydrophilic loops of transendoplasmic reticulum N-acetylglucosamine-1-phosphate transferase[J].Eukaryot Cell,2007,6(2):CR530-535.
[10]Wang JY,Gao CH,Yang YT,et al.An outbreak of the desert sub-type of zoonotic visceral leishmaniasis in Jiashi,Xinjiang Uygur Autonomous Region,People′s Republic of China[J].Parasitol Int,2010,59(3):331-337.
[11]高春花,汪俊云,杨玥涛,等.PCR检测婴儿利什曼原虫无症状感染的研究[J].中国寄生虫学与寄生虫病杂志,2006,24(2):92-96.
[12]Al-Nahhas SA,Kaldas RM.Characterization of Leishmania species isolated from cutaneous human samples from central region of Syria by RFLP analysis[J].ISRN Parasitol,2013,2013:308726.
[13]JirkùM,ZemanováE,Al-Jawabreh A,et al.Development of a direct species-specific PCR assay for differential diagnosis of Leishmania tropica[J].Diagn Microbiol Infect Dis,2006,55(1):75-79.
[14]Spanakos G,Piperaki ET,Menounos PG,et al.Detection and species identification of Old World Leishmania in clinical samples using a PCR-based method[J].Trans R Soc Trop Med Hyg,2008,102(1):46-53.
[15]Saghrouni F,Ga覿ed-Meksi S,Fathallah A,et al.Immunochromatographic r K39 strip test in the serodiagnosis of visceral leishmaniasis in Tunisia[J].Trans R Soc Trop Med Hyg,2009,103(12):1273-1278.
[16]Doudi M,Hejazi SH,Razavi MR,et al.Comparative molecular epidemiology of Leishmania major and Leishmania tropica by PCR-RFLP technique in hyper endemic cities of Isfahan and Bam,Iran[J].Med Sci Monit,2010,16(11):CR530-535.
[17]Bensoussan E,Nasereddin A,Jonas F,et al.Comparison of PCR assays for diagnosis of cutaneous leishmaniasis[J].J Clin Microbiol,2006,44(4):1435-1439.
[18]熊光华,金长发.白蛉栖性及其治理对策的研究进展[J].中国寄生虫学与寄生虫病杂志,2006,24(4):293-298.