Pandey Kishor, Yanagi Testuo, Pandey Basu Dev, Mallik Arun Kumar, Sherchand Jeevan Bahadur, Kanbara Hiroji
Department of Protozoology, Institute of Tropical Medicine, Nagasaki University, Sakamoto 1-12-4, 852-8523 Nagasaki, Japan.
Parasitol Res. 2007 May;100(6):1361-9. doi: 10.1007/s00436-007-0464-4. Epub 2007 Feb 20.
In Nepal, visceral leishmaniasis (VL) is endemic in 13 districts of the central and eastern regions. A total of 166 bone-marrow aspirates were obtained from patients with suspected VL. Ninety-seven were identified as positive by microscopy, and 29 of those were successfully isolated and cultured. We characterized these isolates by molecular analysis and by their ability to infect mice. PCR-restriction fragment length polymorphism analysis of the mini-exon and the cysteine proteinase b gene showed that all isolates were Leishmania donovani, and the restriction pattern of the Nepalese isolates corresponded to the standard Indian strain of L. donovani but differed from that of the Kenyan strain. The single-strand conformation polymorphism analysis of ribosomal internal transcribed spacer showed no genetic heterogeneity within Nepalese isolates. Intraperitoneal inoculation with the promastigotes of all isolates resulted in amastigote proliferation in the spleen of 20 nude mice, of which ten isolates were highly infective, and ten were moderately infective, including one BALB/c mouse. Of the 20 amastigotes isolated from the spleen of nude mice, only the ten highly infective isolates infected BALB/c mice, of which, two isolates were considered to have low infectivity, three isolates were considered to be moderately infective, and five isolates were considered to be highly infective.
在尼泊尔，内脏利什曼病（VL）在中部和东部地区的13个县呈地方性流行。从疑似VL患者身上共采集了166份骨髓抽吸物。其中97份经显微镜检查鉴定为阳性，其中29份成功分离并培养。我们通过分子分析和感染小鼠的能力对这些分离株进行了表征。对小外显子和半胱氨酸蛋白酶b基因的PCR-限制性片段长度多态性分析表明，所有分离株均为杜氏利什曼原虫，尼泊尔分离株的限制性图谱与杜氏利什曼原虫的标准印度株一致，但与肯尼亚株不同。核糖体内部转录间隔区的单链构象多态性分析表明，尼泊尔分离株内无遗传异质性。对所有分离株的前鞭毛体进行腹腔接种，导致20只裸鼠脾脏内无鞭毛体增殖，其中10个分离株感染性高，10个分离株感染性中等，包括1只BALB/c小鼠。从裸鼠脾脏分离出的20个无鞭毛体中，只有10个高感染性分离株能感染BALB/c小鼠，其中2个分离株被认为感染性低，3个分离株被认为感染性中等，5个分离株被认为感染性高。
