2007年第24卷第3期文章目次

摘要:According to the sequence of ATCC VR-2332 strain of porcine reproductive and respiratory syndrome virus published by the Genbank, two pairs of primers were designed for RT-PCR amplification of PRRSV isolates from Liaoning. The corresponding DNA fragments were abtained and , cloned into pMD18-T vector respectively and sequenced in Shanghai. The sequences of ORF567 genes were analysed using DNAStar and compared with those of ATCC VR-2332,CH-1a, BJ-4 LV-M96262 and vaccine strains. Nuclear acid analysis indicates that the homology was up to 89.5%-95% between Liaoning strains and VR-2332,CH-la, B J-4, and 54%-58.3% between Liaoning strains and LV-M96262. Amino acid analysis indicates that the ho- mology was up to 82.8%-94.7% between Liaoning strains and VR-2332,CH-1a,BJ-4 .

猪弓形虫特异性PCR诊断方法的建立及应用

闫若潜 , 刘光辉 , 赵雪丽 , 赵明军 , 盛敏 , 吴志明

2007, 24(3):28-31.

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摘要:A highly sensitive and specific PCR method was developed using the primers designed based on the first internal transcribed spacer ( ITS1) of nuclear ribosomal DNA genome sequences of Toxoplasma gondii (T. gondii) to detect the suspectedly infected pigs. An expected PCR product about 273bp in length was amplified from T. gondii RH strain tachyzoite and blood samples and pulmonary tissues from the pigs with clinical signs. The sensitivity and specificity assays indicated that the PCR method could detect at least 0.001ng DNA of T. gondii RH strain tachyzoite, and no products were amplified from the genomic DNA of the other 9 species of pathogenic microorganism. The PCR was used to amplify the DNA of 30 samples from suspiciously infected pigs and 60 samples from healthy pigs.And 24 from 30 suspicious samples as well as 5 from 60 healthy samples showed positive reactions. Two PCR products amplified from suspicious samples were randomly picked out and sequenced, and the homology ratios of nucleotide and amino acids sequences of the two PCR products with the corresponding region of published T. gondii genome in GenBank were both 100%. respectively, suggesting that the established PCR method was highly specific and sensitive,and was suitable to clinic rapid T. gondii diagnosis.

新城疫病毒TL1株P基因的克隆及序列分析

王学理 , 王兴龙 , 李晓艳 , 任林柱 , 张辉 , 闫广谋