福建猫源弓形虫IgG抗体检测及PCR-RFLP基因分型与遗传变异分析

Detection of antibody IgG,PCR-RFLP genotyping and sequence variation analysis of Toxoplasma gondii from cat in Fujian

采用酶联免疫吸附(ELISA)试验对福建某花鸟市场采集的40份猫血液样本进行弓形虫IgG抗体检测,对抗体呈现阳性的血液样品进行弓形虫B1基因PCR扩增;对扩增结果呈现阳性的样本采用多位点巢式PCR-RFLP技术,在SAG1、5’SAG2、3’SAG2、Alt.SAG2、SAG3、BTUB、GRA6、c22-8、c29-2、L358、PK1和Api co共12个位点进行弓形虫基因分型研究和部分位点的遗传变异分析。结果表明,40份血清中,19份血清呈弓形虫IgG抗体阳性,阳性率为47.5%,19份血清抗体呈阳性的血液样品中,B1基因PCR扩增出3份阳性样品,进一步多位点巢式PCR-RFLP分析和遗传变异分析发现,PCR阳性样品中的弓形虫基因型均表现为New Genotype型。

The IgG against Toxoplasma gondii was detected for 40 cat blood samples from a pet market in Fujian using ELISA method and then T. gondii DNAfrom the positive blood samples was tested by PCR using T. gondii B1 gene primers. The genetic diversity of T. gondii DNA was typed at 12 gentic markers using the multilocus PCR-RFLP,including nuclear loci SiGl,5＇SiG2,3＇SAG2,alternative SAG2, SAG3, BTUB,GRA6,c22-8,c29-2,L358,PK1 and an apicoplast lucus Apico. The sequences of SAG1,SAG2 and SAG3 were used for variation analysis. The results showed that the positive rate of IgG against T. gondii was 47.5%（19/40） in cat blood samples. Three T. gondii B1 gene DNiwas detected by PCR amplification in these 19 positive blood samples. The mu]tilocus PCR-RFLP results and the sequences of 3 loci were analyzed and compared with reference strains RH（Type Ⅰ ）,PRU（TypeⅡ ）,and the result revealed the 3 gondiiisolatesinFujian were found tobeanunique genotype and groupedtoanewgenotypeinthisstudy.