微卫星DNA在动物亲子鉴定中的应用及研究进展

本文概述了微卫星DNA进行亲子鉴定的基本原理方法，将其用于亲子鉴定的优点以及研究进展，最后对未来的应用进行了展望。     

微卫星DNA在牛亲子鉴定方面的研究进展

微卫星DNA因具有长度多态性这一特点,而被广泛应用于个体的亲子鉴定。作者概述了微卫星DNA在牛亲子鉴定方面的研究进展及发展趋势。     

DNA生物传感器在疾病检测中的研究进展和应用前景

DNA生物传感器的近年发展起来的一种新型生物传感器，它可以从基因水平上对引起疾病的病原作出定性、定量的检测，其简单、快速和准确的特点优越于传统的诸多分析手段，因此DNA生物传感器在疾病的临床检验、基因诊断上有极大的应用前景。     

进口基因检测用动植物DNA/RNA风险分析

近年来,我国在基因检测方面正在积极抢占国际市场份额。然而该产业的时效性要求很高,因此进出口通关效率至关重要。北京出入境检验检疫局在对进口基因检测用动植物DNA/RNA进行系统地风险分析后认为,经纯化的动植物DNA/RNA不具备感染其它生物体的条件,无传染性,检疫风险极低,并据此提出了检疫监管建议。     

应用线粒体DNA鉴定动物源性饲料成分研究进展

为避免牛海绵状脑病和羊痒病疫区的蛋白饲料进口造成畜牧业上的经济损失,世界各国纷纷开展用分子生物学方法检测牛、羊源性蛋白饲料,本文就运用分子生物学方法有效检测动物物种间线粒体DNA特异性以确认动物源性蛋白饲料成分的来源作出阐述。     

建设省级示范畜禽疾病检测实验实训中心的实践与思考

安徽农业大学为适应动物医学和动物检疫学发展的需要,深化实践教学改革,整合校内临床检测实验室与社会实践基地,建设成省级示范畜禽疾病检测实验实训中心。该中心通过优化整合实验实训教学内容,强化实践教学环节,构建创新实验实训教学体系,提高临床教学水平,充分发挥培养应用型动物医学和动物检疫学专门人才以及服务社会的功能。     

免疫胶体金技术基本原理及其在疾病检测中的应用进展

近年来胶体金标记技术迅速发展,应用范围也逐渐扩大,不仅应用于光镜和电镜水平定位、定性及定量研究含有抗原物质的组织、细胞及亚细胞结构,还应用于免疫转印、流式细胞术、液相免疫测定、固相斑点金/银染色、免疫层析法(immunochromatography)和快速免疫金渗滤法(Dot immuogol     

动物产地检疫工作的调查与思考

动物产地检疫是指动物检疫人员在动物离开饲养生产地之前所进行的检疫,也就是我们通常说的到场、到户、到指定地点的检疫,是动物检疫监督的重要环节和第一道关口。随着社会经济迅速发展,动物及动物产品流通日益频繁,动物产地检疫已经日益显示出极为重要的作用。     

动物检疫中检测试剂存在的问题及对策

开展动物检疫工作会在一定程度上保证人们的身体健康,特异、敏感的检测试剂是顺利完成检疫工作的保障.本文主要提出了我国动物检疫中检测试剂遇到的问题,并提出相应的措施,为我国动物检疫选用试剂提供技术支撑.     

AFLP fingerprinting for paternity testing in ducks

1. The accuracy and reproducibility of AFLP fingerprinting was investigated in the duck (Anas Platyrhynchos), using a multicolour fluorescent labeling technique. The fluorescent labelling fragments were separated on a capillary electrophoresis-base ABI PRISM 3100 Genetic Analyzer. 2. A total of 337 AFLP peaks with 103 of them being polymorphic markers were generated by 16 sets consisting of EcoRI/TaqI primer pair combinations. The number and size range of AFLP polymorphisms detected per primer pair varied from 3 to 11 and 58 to 290 bp, respectively. About 30.6% (103/337) of AFLP peaks were detected polymorphisms, with an average of 6.4 polymorphic markers per primer pair. 3. The clear polymorphic peaks were amplified with EcoR+AC/Taq+AC primer combinations. The AFLP peaks showed high reproducibility. From the family testing, we found that the fingerprints of all the offspring were derived from one or other parent. Therefore, we conclude that AFLP fingerprinting might be a suitable method for duck paternity testing.     

An unusual case from paternity testing in dogs

We are solving disputed paternities in purebred dogs using microsatellite analysis. An interesting case involved four puppies for which pedigrees were already issued by the Swiss Kennel Club. The investigation had to be carried out without a sample from the stated sire. Even though the relationship of this sire could not be determined conclusively, comparison of microsatellite alleles within this family, including an alleged half-brother of the four puppies, showed clearly that the breeder had made contradictory statements and that false recordings of matings had occurred. The pedigrees for these four puppies were withdrawn and sanctions against the breeder were imposed.     

利用微卫星标记进行猪亲子鉴定

以杜洛克猪、长白猪、大白猪3个品种的48头系谱清楚的猪为样本,使用PAGE法从55个微卫星位点中筛选出扩增效果好、多态性丰富且便于进行多重PCR扩增的位点,对试验样本逐一进行基因分型,并据此进行亲权确认,建立一种高效的亲子鉴定方法。结果显示,试验选取的14个微卫星位点在48个样本中进行排父率分析,在双亲未知、只知单亲和双亲已知的情况下计算的累积排父率分别为0.978 5,0.998 4,0.999 99;利用建立的亲子鉴定体系对样本中8个家系进行检测,结果累积排父率均大于99.99%。结果表明,试验构建的微卫星标记体系,为亲本信息不明的个体进行亲子关系的验证,纠正潜在的错误系谱,建立完整、准确的系谱体系,保证育种值估计的准确性,保障猪育种工作的正常开展具有非常重要的意义。     

Analysis of microsatellites and paternity testing in Rasa Aragonesa sheep

Four microsatellite loci (MAF50, MAF18, OarFCB20 and MCM527) were studied in Rasa Aragonesa sheep in order to evaluate their use in paternity testing. Several population characteristics were estimated [allele frequencies. effective allele number (Ne), polymorphism informative content (PIC) and probability of excluding wrong paternities (Pe)]. In 32 randomly chosen individuals, four alleles were detected for MAF50, with 2.55 effective alleles, 0.58 PIC and 0.35 Pe. For MAF18, five alleles were identified, with 2.99 effective alleles, 0.51 PIC and 0.32 Pe. For oarFCB20, 10 alleles were observed, with 6.06 effective alleles, 0.82 PIC and 0.68 Pe. Finally, for MCM527, six alleles were found, with 3.75 effective alleles, 0.69 PIC and 0.50 Pe. When these loci were used together with serum transferrin locus, Pe rose to 97.20 per cent. Field trials confirmed the real usefulness of these techniques.     

Molecular and cytogenetic paternity testing of a male offspring of a hinny

An alleged male foal of a female mule, whose sire and grandparents were unknown, was identified for its pedigree. Parentage testing was conducted by comparing polymorphism of 12 microsatellite DNA sites and mitochondrial D‐loop sequences of the male foal and the female mule. Both the sequence analysis of species‐specific DNA fragments and a cytogenetic analysis were performed to identify the species of the foal and its parents. The results showed that the alleged female mule is actually a hinny, and the male foal, which possesses 62 chromosomes, qualifies as an offspring of the female hinny and a jack donkey.     

Performance of using opposing homozygotes for paternity testing in Japanese Black cattle

Genome-wide single nucleotide polymorphism (SNP) markers in Japanese Black cattle enable genomic prediction and verifying parent–offspring relationships. We assessed the performance of opposing homozygotes (OH) for paternity testing in Japanese Black cattle, using SNP genotype information of 50 sires and 3,420 fattened animals, 1,945 of which were fathered by the 50 genotyped sires. The number of OH was counted for each sire–progeny pair in 28,764 SNPs with minor allele frequencies of ≥0.05 in this population. Across all pairs of animals, the number of OH tended to increase as the pedigree-based coefficient of relationship decreased. With a threshold of 288 (1% of SNPs) for paternity testing, most sire–progeny pairs were detected as true relationships. The frequency of Mendelian inconsistencies was 2.4%, reflecting the high accuracy of pedigree information in Japanese Black cattle population. The results indicate the utility of OH for paternity testing in Japanese Black cattle.     

The value of DNA paternity identification in beef cattle: examples from Nevada's free-range ranches

The feasibility and economic value of DNA paternity identification were investigated and illustrated using Nevada beef cattle operations. A panel of 15 microsatellites was genotyped in 2,196 animals from 8 ranches with a total of 31,571 genotypes. Probabilities of exclusion for each marker within ranch and across ranches were computed. Joint probabilities of exclusion for the 15 microsatellites were also determined, resulting in values over 0.99 for any individual ranch and across ranches. Dropping 1 or 2 microsatellites with the lowest probabilities of exclusion resulted in joint probabilities greater than 0.99 and with marginal reduction compared with the probabilities with 15 microsatellites. Formulas for benefit-cost analysis for a DNA paternity identification program in beef cattle were derived. Genotyping 15 microsatellites with 20 calves per sire resulted in benefits of $1.71 and $2.44 per dollar invested at bull culling rates of 0.20 and 0.30, respectively. The breakpoints for the program to be profitable occurred when the ratio of the price of 1 kg of calf liveweight over the cost of genotyping 1 microsatellite was greater than 1.1 for a bull culling rate of 0.30. Benefit-cost analysis was also derived under incomplete DNA paternity identification using a lower number of DNA markers than necessary to achieve joint probabilities of exclusion of 0.99. Approximately a 20% increase in the benefit-cost ratio was achieved using 10 vs. 12 microsatellites with incomplete paternity identification. The greater the number of bulls in the operation, the lower the benefit-cost ratio of the paternity testing program. Low probabilities of exclusion and a high number of bulls in the beef operation reduced the benefit-cost ratio dramatically. The DNA paternity identification programs are feasible and may be profitable for free-range beef cattle operations.     

Use of DNA bar codes to resolve a canine paternity dispute

The DNA fingerprinting method was used to resolve a canine paternity dispute. During the same estrus, a Shih Tzu bitch was inseminated by 2 dogs--a Shih Tzu and a Coton de Tulear. Because both breeds are alike phenotypically, it was difficult to decide whether the pups were purebred or of mixed breeding. The DNA bar codes indicated unambiguously that the 2 sires had fathered one pup each, thus documenting superfecundation.     

Use of DNA fingerprinting in paternity analysis of closely-related Exmoor ponies

DNA fingerprinting techniques were used to try to resolve the parentage of an Exmoor pony foal. Three young Exmoor ponies, one female and two males, shared a paddock and the female subsequently became pregnant. The two possible sires were three-quarter siblings and were also half-siblings to the dam. Southern hybridisation of Exmoor pony DNA with human mini-satellite probes resolved the disputed parentage in spite of the fact that there was a 70 per cent band share between the individuals involved. Colt M6 was 2.06 times more likely to be the father than an uncle, and Colt M3 was 477 times more likely to be an uncle than the father.