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一种检测鉴定昆虫体内Wolbachia感染的特异性引物、方法和试剂盒

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摘要

本发明公开了一种检测鉴定昆虫体内Wolbachia感染的特异性引物、方法和试剂盒。所述特异性扩增引物Tw16s1，包括正向引物F和反向引物R，其序列依次如SEQ ID NO：1～2所示。所述检测方法为：S1.提取待测昆虫总DNA；S2.以S1的DNA为模板，利用所述引物进行PCR扩增反应；S3.将S2的PCR扩增反应产物电泳检测，从而进行判断。本发明所述引物特异性强，扩增产物为单一目的片段，而且灵敏度高，能筛选出假阴性、非特异性的样品，从而提高检出率。从提DNA到获得结果仅需要4个小时左右。可以精确、快速地发现检测出样品中是否受Wolbachia感染，为进一步得用进行疾病预防和对昆虫进行生殖调控提供基础。

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公开/公告号CN108707681B

专利类型发明专利
公开/公告日2022-02-18

原文格式PDF
申请/专利权人华南农业大学;
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申请/专利号CN201810504166.5
发明设计人何晓芳;林良冠;张美娇;
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申请日2018-05-23
分类号C12Q1/689(20180101);C12Q1/686(20180101);C12Q1/04(20060101);C12N15/11(20060101);
代理机构44102 广州粤高专利商标代理有限公司;
代理人林丽明
地址 510642 广东省广州市天河区五山路483号
入库时间 2022-08-23 13:08:22

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1. 一种检测鉴定昆虫体内Wolbachia感染的特异性引物、方法和试剂盒 [P] . 中国专利： CN108707681B . 2022.02.18
2. 一种检测鉴定昆虫体内Wolbachia感染的特异性引物、方法和试剂盒 [P] . 中国专利： CN108707681A . 2018-10-26
3. Processes for detecting a mutation in the nucleic acid to detect fungal and fungal resistance to an analogue of estobilurina,Fungal DNA sequence encoding all or part of a proteu00ecna of cytochrome b, cytochrome b proteu00ecna fungal.The allele specific oligonucleotu00ecdeo capable of connecting to a sequence of fungal nucleic acid, initiator of diagnosis or diagnostic oligonucleotu00ecdeo capable of connecting to a template.One or more diagnostic primers, probe the allele specific oligonucleotu00ecdeo, diagnostic kit to detect fungal pathogen resistance, process plant, a fungicide.To detect and quantify the frequency of mutation, to select a fungicide active and their nu00ecveis optimal implementation, and to control the fungal infection of a collection, and,A computer legu00ecvel [P] . BR0010163A . 2002-01-15

机译：用于检测核酸突变的方法，以检测对食管胆囊炎类似物的真菌和真菌抗性，真菌DNA序列编码全部或部分细胞色素b蛋白，细胞色素b蛋白真菌，等位基因特异性寡核苷酸能够连接到真菌核酸序列的诊断或诊断寡核苷酸的启动子，能够连接到模板。一个或多个诊断引物，探查等位基因特异性寡核苷酸，诊断试剂盒以检测真菌病原体抗性，加工厂要检测和量化突变的频率，选择一种活性杀菌剂及其最佳实施方案，控制一个集合体的真菌感染以及计算机支路，
4. Processes for the identification of an elite event in biological samples, for the confirmation of the purity of seed, for the selection of seeds in the presence of said event elite.For the determination of the status of zigosidade of a plant, material of plant or seed which comprises the said event elite, to detect the presence of said event elite.And for the production of a plant or a seed of cotton which comprises the said event elite, as well as kits for the identification of said event elite.A pair of primers suitable for use in specific detection of the same, and specific probe for the identification of said event elite [P] . BRPI0614338B1 . 2018-01-02

机译：用于鉴定生物样品中优良事件，确认种子纯度，在存在所述优良事件的情况下选择种子的方法。用于确定植物，植物或植物的齐哥状态的状态包括所述事件精英的种子，以检测所述事件精英的存在。并且用于生产包含所述事件精英的植物或棉花种子，以及用于鉴定所述事件精英的试剂盒。适用于特异性检测的引物的鉴定以及用于鉴定所述事件精英的特异性探针
5. Control primer annealing to improve the specificity of annealing in nucleic acid amplification kit.Methods for amplifying a nucleic acid sequence of a DNA or a mixture of nucleic acids.And for selectively amplifying a nucleic acid sequence of a target DNA or a mixture of nucleic acids, and a sequence of the target nucleic acid from a mRNA.Method for detecting the complementarity of DNA to mRNA differentially expressed in two or more samples of nucleic acid, methods for amplifying a fragment of the cDNA and DNA quickly target.And to amplify a population of cDNAs of filament double full-length complementary to mRNAs cDNAs, and filament double fortified with 5 'complementary to mRNAs,Method to amplify rather than a sequence of nucleotu00ecdeo target at the same time, methods for producing a digital printing gDNA DNA, and RNA in a sample of mRNA.Method for identifying segments of homology is conserved in a family of multigene from a sample of mRNA, a method for the mutagenesis in a target [P] . BR0214741A . 2004-11-23

机译：控制引物退火，以提高核酸扩增试剂盒中退火的特异性。扩增DNA或核酸混合物的核酸序列的方法以及选择性扩增靶DNA或核酸混合物的核酸序列的方法用于检测DNA与两个或多个核酸样品中差异表达的mRNA的互补性的方法，扩增cDNA和DNA片段的方法可快速靶向。全长与mRNA cDNA互补的双细丝cDNA的cDNA群体，以及与mRNA互补的5'增强双细丝的cDNA，同时扩增而不是核苷酸靶序列的方法，产生数字印刷gDNA DNA的方法mRNA样品中的RNA和RNA.mRNA样品的多基因家族中用于鉴定同源片段的方法是保守的，这是一种在靶标中诱变的方法