Maleki-Ravasan N, Oshaghi Ma, Javadian E, Rassi Y, Sadraei J, Mohtarami F
Department of Medical Parasitology and Entomology, College of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran J Arthropod Borne Dis. 2009;3(1):8-18. Epub 2009 Jun 30.
We aimed to develop a PCR-RFLP assay based on available sequences of putative vertebrate hosts to identify blood meals ingested by field female sand fly in the northwest of Iran. In addition, the utility of PCR-RFLP was compared with ELISA as a standard method.
This experimental study was performed in the Insect Molecular Biology Laboratory of School of Public Health, Tehran University of Medical Sciences, Iran in 2006-2007. For PCR-RFLP a set of conserved vertebrate primers were used to amplify a part of the host mitochondrial cytochrome b (cyt b) gene followed by digestion of the PCR products by Hae III enzyme.
The PCR-RFLP and ELISA assays revealed that 34% and 27% of field-collected sand flies had fed on humans, respectively. Additionally, PCR-RFLP assays could reveal specific host DNA as well as the components of mixed blood meals. Results of PCR-RFLP assay showed that the sand flies had fed on cow (54%), human (10%), dog (4%), human and cow (21%), dog and cow (14%), and human and dog (3%).
The results can provide a novel method for rapid diagnosis of blood meal taken by sandflies. The advantages and limitations of PCR and ELISA assays are discussed.
我们旨在基于假定脊椎动物宿主的现有序列开发一种聚合酶链反应-限制性片段长度多态性（PCR-RFLP）检测方法，以鉴定伊朗西北部野外雌性白蛉摄取的血餐。此外，将PCR-RFLP的效用与作为标准方法的酶联免疫吸附测定（ELISA）进行了比较。
这项实验研究于2006 - 2007年在伊朗德黑兰医科大学公共卫生学院昆虫分子生物学实验室进行。对于PCR-RFLP，使用一组保守的脊椎动物引物扩增宿主线粒体细胞色素b（cyt b）基因的一部分，然后用Hae III酶消化PCR产物。
PCR-RFLP和ELISA检测显示，分别有34%和27%的野外采集的白蛉吸食过人类血液。此外，PCR-RFLP检测能够揭示特定的宿主DNA以及混合血餐的成分。PCR-RFLP检测结果表明，白蛉吸食过牛血（54%）、人血（10%）、狗血（4%）、人血和牛血（21%）、狗血和牛血（14%）以及人血和狗血（3%）。
这些结果可为快速诊断白蛉摄取的血餐提供一种新方法。讨论了PCR和ELISA检测的优缺点。
