利用聚合酶链式反应（PCR）检测法对取自皮肤利什曼病患者的吉姆萨染色玻片进行检测，以鉴定利什曼原虫的种类。
Identification of leishmania species using PCR assay on giemsa-stained slides prepared from cutaneous leishmaniasis patients.
作者信息
Kheirandish Farnaz, Chegeni Sharafi Ali, Kazemi Bahram, Mohebali Mehdi, Sarlak Amanollah, Tarahi Mohamad Javad, Holakouee Kourosh, Hajaran Homa
机构信息
Razi Herbal Medicines Research Center, Department of Parasitology and Mycology, Lorestan University of Medical Sciences, Khorramabad, Iran.
Department of Communicable Disease Control and Prevention, Deputy of Health, Lorestan University of Medical Sciences, Khorramabad, Iran.
出版信息
Iran J Parasitol. 2013 Jul;8(3):382-8.
BACKGROUND
Leishmaniasis is a group of diseases that are created by intracellular parasites of Leishmania. Cutaneous leishmaniasis is considered as one of the health problems in some provinces of Iran.
METHODS
In this study, a total of 178 Giemsa-stained slides from confirmed cases of cutaneous leishmaniasis were examined. The slides were prepared from the patients with cutaneous leishmaniasis that referred to health centers and infected during the epidemic of cutaneous leishmaniasis in Poldokhtar city, Lorestan Province, Iran in 2006.Genomic DNA from each slide was extracted. After DNA extraction, ITS-PCR was used.
RESULTS
Out of 178 slides, 129 (72.47%) samples had a band in the range of 485 bp and 49 (27.53%) samples 626 bp that matched L. tropica and L. major standard samples, respectively.
CONCLUSION
This study showed that Leishmania DNA could be efficiently extracted and amplified even from old Giemsa-stained microscopic slides that were stored more than 6 yr. In this study was shown that both L. tropica and L. major species exist in Lorestan Province.
背景
利什曼病是由利什曼原虫细胞内寄生虫引发的一组疾病。皮肤利什曼病被视为伊朗某些省份的健康问题之一。
方法
在本研究中，共检查了178张来自确诊皮肤利什曼病病例的吉姆萨染色玻片。这些玻片取自2006年转诊至伊朗洛雷斯坦省波尔多克塔尔市卫生中心且在皮肤利什曼病流行期间感染的皮肤利什曼病患者。从每张玻片提取基因组DNA。DNA提取后，采用ITS-PCR。
结果
在178张玻片中，129份（72.47%）样本在485 bp范围内有条带，49份（27.53%）样本在626 bp处有条带，分别与热带利什曼原虫和硕大利什曼原虫标准样本匹配。
结论
本研究表明，即使从保存超过6年的陈旧吉姆萨染色显微玻片中也能有效提取和扩增利什曼原虫DNA。本研究表明洛雷斯坦省同时存在热带利什曼原虫和硕大利什曼原虫这两种物种。
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