Lah Ernieenor Faraliana Che, Ahamad Mariana, Haron Mohd Subail, Ming Ho Tze
Acarology Unit, Infectious Diseases Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia.
Asian Pac J Trop Biomed. 2012 Mar;2(3):223-7. doi: 10.1016/S2221-1691(12)60046-X.
To establish a polymerase chain reaction (PCR) technique based on cytochrome b (cytb) gene of mitochondria DNA (mtDNA) for blood meal identification.
The PCR technique was established based on published information and validated using blood sample of laboratory animals of which their whole gene sequences are available in GenBank. PCR was next performed to compile gene sequences of different species of wild rodents. The primers used were complementary to the conserved region of the cytb gene of vertebrate's mtDNA. A total of 100 blood samples, both from laboratory animals and wild rodents were collected and analyzed. The obtained unknown sequences were compared with those in the GenBank database using BLAST program to identify the vertebrate animal species.
Gene sequences of 11 species of wild animals caught in 9 localities of Peninsular Malaysia were compiled using the established PCR. The animals involved were Rattus (rattus) tanezumi, Rattus tiomanicus, Leopoldamys sabanus, Tupaia glis, Tupaia minor, Niviventor cremoriventor, Rhinosciurus laticaudatus, Callosciurus caniseps, Sundamys muelleri, Rattus rajah and Maxomys whiteheadi. The BLAST results confirmed the host with exact or nearly exact matches (>89% identity). Ten new gene sequences have been deposited in GenBank database since September 2010.
This study indicates that the PCR direct sequencing system using universal primer sets for vertebrate cytb gene is a promising technique for blood meal identification.
建立一种基于线粒体DNA（mtDNA）细胞色素b（cytb）基因的聚合酶链反应（PCR）技术用于血餐鉴定。
根据已发表的信息建立PCR技术，并使用其全基因序列可在GenBank中获取的实验动物血样进行验证。接下来进行PCR以编译不同种类野生啮齿动物的基因序列。所用引物与脊椎动物mtDNA的cytb基因保守区域互补。共收集并分析了100份来自实验动物和野生啮齿动物的血样。使用BLAST程序将获得的未知序列与GenBank数据库中的序列进行比较，以鉴定脊椎动物物种。
使用建立的PCR编译了在马来西亚半岛9个地点捕获的11种野生动物的基因序列。涉及的动物有黑家鼠、蒂奥曼鼠、萨氏猪尾鼠、树鼩、小鼩鼱、白腹鼠、大赤鼯鼠、犬头松鼠、穆氏巨鼠、拉贾鼠和白氏东方鼠。BLAST结果证实宿主与精确或几乎精确匹配（>89%同一性）。自2010年9月以来，已有10条新的基因序列存入GenBank数据库。
本研究表明，使用通用引物组对脊椎动物cytb基因进行PCR直接测序系统是一种有前途的血餐鉴定技术。
