[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put...[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put into capsule was fed to chickens. At the same time, the collected oocysts were identified by PCR method. [Result] The oocysts were isolated from feces of 15 chickens among that of 20 chickens and the infection rate was 75%. The PCR results demonstrated that the single-oocyst strain was E.tenella. [Conclusion] The inoculation of single oocyst capsule was simple, besides, this method did not only save time but also declined inoculation difficulty, increased infection rate and provided good materials for biological research of coccidian.展开更多
In order to develop a more applicable DNA molecular marker identification method for theauthentication of Bungarus parvus and its adulterants,DNA templates were extracted from Bungarus parvus.adulterants of the crude ...In order to develop a more applicable DNA molecular marker identification method for theauthentication of Bungarus parvus and its adulterants,DNA templates were extracted from Bungarus parvus.adulterants of the crude drugs and their original animals.Cyt b gene fragment was amplified from all these templatesrespectively using universal primers.PCR products were purified and sequenced directly.After sequence alignmentand comparing the sequence of Bungarus parvus to its adulterants.we find that Cyt b gene is a good molecularmarker for authentication of the crude drugs since interspecific differentiation is more evident than that ofintraspecies in DNA sequence of the gene.Based on the sequence data of Cyt b gene fragment,a pair of highlyspecialized primers,used as diagnostic primers,was designed for the PCR identification of the crude drugs.Usingdiagnostic primers for the PCR identification,Bungarus parvus samples could be absolutely distinguished from all itsadulterants when annealing temperature is at 60°℃~65°℃,and no incorrect or missing discrimination was found underthese PCR conditions.Mixed powder of the qualified crude drug and its adulterants could also be detected bydiagnostic PCR.The results indicate that diagnostic PCR using the primers designed in the present study is a simple,effective and applicable method for the authentication of Bungarus parvus,and this method might also be a new wayfor examining the compositions of Chinese patent medicine.展开更多
基金Supported by the National Natural Science Foundation of China(30671580, 30170696)~~
文摘[Objective] In order to get a purified strain to carry out the relative molecular biology research about E.tenella. [Method] The single-oocyst isolation method was improved and the isolated single-oocyst which was put into capsule was fed to chickens. At the same time, the collected oocysts were identified by PCR method. [Result] The oocysts were isolated from feces of 15 chickens among that of 20 chickens and the infection rate was 75%. The PCR results demonstrated that the single-oocyst strain was E.tenella. [Conclusion] The inoculation of single oocyst capsule was simple, besides, this method did not only save time but also declined inoculation difficulty, increased infection rate and provided good materials for biological research of coccidian.
基金Supported by NSFC No.39570865,No.39870913grant from Jiangsu Educational Committee(No.98KJB360010).
文摘In order to develop a more applicable DNA molecular marker identification method for theauthentication of Bungarus parvus and its adulterants,DNA templates were extracted from Bungarus parvus.adulterants of the crude drugs and their original animals.Cyt b gene fragment was amplified from all these templatesrespectively using universal primers.PCR products were purified and sequenced directly.After sequence alignmentand comparing the sequence of Bungarus parvus to its adulterants.we find that Cyt b gene is a good molecularmarker for authentication of the crude drugs since interspecific differentiation is more evident than that ofintraspecies in DNA sequence of the gene.Based on the sequence data of Cyt b gene fragment,a pair of highlyspecialized primers,used as diagnostic primers,was designed for the PCR identification of the crude drugs.Usingdiagnostic primers for the PCR identification,Bungarus parvus samples could be absolutely distinguished from all itsadulterants when annealing temperature is at 60°℃~65°℃,and no incorrect or missing discrimination was found underthese PCR conditions.Mixed powder of the qualified crude drug and its adulterants could also be detected bydiagnostic PCR.The results indicate that diagnostic PCR using the primers designed in the present study is a simple,effective and applicable method for the authentication of Bungarus parvus,and this method might also be a new wayfor examining the compositions of Chinese patent medicine.
