Improvement and application of AP-PCR identification for Leismania parasites

莱克曼原虫AP-PCR鉴定方法的改进及应用

• 批准号: 12670235
• 负责人: MIMORI Tatsuyuki
• 金额: $ 2.18万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670235/
• 关键词: Leishmania parasite; Identification; PS-PCR; Diagnosis; Sampling; Cotton swab; Sandfly; Epidemiology; 皮膚型リーシュマニア症

Leishmaniasis caused by the genus Leishmania parasites are a group of diseases having a worldwide distribution in the tropical and temperate zones. Approximately 12 million people in 88 countries around the world are already affected by the diseases where they present a considerable health problem. In our country, patients of leishmaniasis were found as imported disease. The precise identification of Leishmania species is important for the appropriate treatment regimen and public health surveillance since different species cause different clinical features of the disease. Polymerase chain reaction (PCR) methods are used to identify the Leishmania parasites. Recently, we have also developed a highly specific PCR panel to enable the identification of the five major Leishmania species which cause NWCL by polymorphism-specific (PS) -PCR diagnosis (Mimori et al, 1998) were performed with formalin-fixed biopsy specimens of the leishmanial lesions-However, it is not so easy to take biopsy mat … More erials from the patients in the field level, using punch under local anesthesia. It is still necessary to search for a better sampling method to get materials for PCR examination, avoiding patients suffering from biopsy injury. PCR for diagnosis of cutaneous leishmaniasis was compared using different samples such as exudate, syringe-sucked fluid and biopsy specimens taken from ulcerative lesions of patients. The sensitivity of PCR was same when exudates and syringe-sucked fluid samples from the skin lesions as well as biopsy materials were used, considering parasitological diagnosis (smear and culture) as gold standard. Moreover, we performed the easy sampleing method of exudates using cotton swab on the field examination in an endemic area of Ecuador. Considering the collection method of non-injurious to the patients it can be suggested that the exudate samples using cotton swab for the diagnosis of cutaneous leishmaniasis by PCR would be a better than biopsy samples. It was performed that the study were find out the infection rate of the sandflies by Leishmania parasite and to know the usefulness of PCR to detect Leishmania species from sandfly. PCR may be also a useful tool to detect Leishmania species from infected sandflies and thus may be used for future epidemiological surveys in leishmaniasis endemic areas. Less

由利什曼原虫属寄生虫引起的利什曼病是一组在热带和温带地区具有世界性分布的疾病。全世界88个国家中约有1 200万人已经受到这些疾病的影响，这些疾病构成了相当大的健康问题。在我国，利什曼病患者多为输入性疾病。准确鉴定利什曼原虫种类对于适当的治疗方案和公共卫生监测非常重要，因为不同的种类导致疾病的不同临床特征。聚合酶链反应（PCR）方法用于鉴定利什曼原虫。最近，我们还开发了一种高度特异性的PCR板，以通过多态性特异性（PS）鉴定引起NWCL的五种主要利什曼原虫。PCR诊断（Mimori et al，1998）是用福尔马林固定的利什曼原虫病变的活检标本进行的。 ...更多信息 在局部麻醉下使用打孔器，从现场的病人那里采集病历。仍需寻找更好的取材方法，以获得PCR检测材料，避免患者遭受活检损伤。比较了皮肤利什曼病患者溃疡性病变渗出液、抽吸液及活检标本的PCR诊断结果。以寄生虫学诊断（涂片和培养）为金标准，当使用皮损渗出液和分泌物抽吸液样本以及活检材料时，PCR的灵敏度相同。此外，我们在厄瓜多尔的一个流行区进行了现场检查的渗出液用棉拭子的简单采样方法。考虑到对患者无损伤的采集方法，可以建议使用棉拭子采集渗出液样本用于PCR诊断皮肤利什曼病将优于活检样本。本研究旨在了解白蛉体内利什曼原虫的感染率，了解PCR技术在白蛉体内利什曼原虫检测中的作用。PCR也可能是一个有用的工具，检测感染白蛉利什曼原虫物种，因此可能用于未来的流行病学调查在利什曼病流行区。少

项目成果

H.Uezato et al.: "Detection of species of the subgenus Leishmania parasites using polymerase chain reaction and southern Blotting"J. Dermatol.. 28. 475-480 (2001)

H.Uezato 等人：“利用聚合酶链式反应和 Southern Blotting 检测利什曼原虫亚属寄生虫的种类”J.

M.H.Calvopina et al.: "Leishmaniasis en Ecuador : diagnostico de la leishmaniasis cetanea con la reaccion en cadena de la polimerasa (PCR) en comparacion con las tecnnicas convencionales."Educa.Med.Continu.. 66. 14-21 (2000)

M.H.Calvopina 等人：“厄瓜多尔利什曼病：诊断利什曼病 cetanea con la reaccion en cadena de la polimerasa (PCR) en comparacion con las tecnnicas conventionionales.”Educa.Med.Continu.. 66. 14-21 (2000)

M.H.Calvopina: "Leishmaniasis en Ecuador : Diagnostico de la leishmaniasis cutanea con la reaccion en cadena de la polimerasa (PCR) en comparacion con las tecnicas convencionales"Educa. Med. Continu.. 66. 121-131 (2000)

M.H.Calvopina：“厄瓜多尔利什曼病：皮肤利什曼病诊断与聚合酶链反应 (PCR) 与常规技术比较”教育。

Y.Hashiguchi: "Estudios soble la Leishmaniasis en el Nuevo Mundo y su Transmision, con Especial Referencia al Ecuador. No. 5"Kyowa Printing (Kochi, Japan). 193 (2000)

Y.Hashiguchi：“Estudios soble la Leishmaniasis en el Nuevo Mundo y su Transmission，con Especial Referencia al Ecuador。No. 5”Kyowa Printing（日本高知）。

Y. Hashiguci ed: "Studies on New World leishmaniasis and its transmission, with particular reference to Ecuador. No.6"Kyowa Printing (Kochi, Japan). 218 (2001)

Y. Hashiguci 编辑：“新世界利什曼病及其传播研究，特别是厄瓜多尔。第 6 期”Kyowa Printing（日本高知）。