The control and prevention of pasteurellosis in rabbits which makes the hosts unsuitable for experimental use raised the needs to improve and simplify the procedures of Enzyme-Linked Immunosorbent Assay (ELISA) for de...The control and prevention of pasteurellosis in rabbits which makes the hosts unsuitable for experimental use raised the needs to improve and simplify the procedures of Enzyme-Linked Immunosorbent Assay (ELISA) for detect of antibody against P.multocida. A comparison on the sensitivity and specificity of bacterial culture of antemortem and postmortem samples, complement fixation test and enzyme-linked immunosorbent assay of 11 apparently healthy adult rabbits was conducted. The incidence rates showed 45.45%,54.54% and 72.73% respectively. The sensitivity for the three methods were 0.63, 0.67,and 1.00,and specificity for them were 1.00, 0.67 and 1.00 respectively. Somatic serotypes of isolates of P.multocida from rabbits of three groups (rabbits of group 2 were with clinic signs, those of groups 1 and 3 were apparently healthy) revealed no remarkable differences,and the predominant types were type 3 and type 3, 4. This was somewhat different from the reports derived from other states. As the antigen of different serotype used in ELISA may have different sensitivity and specificity, which is affected also by different preparation method, a type non-specific antigen should be selected to meet such request. The trial of accomplishment of ELISA without positive and negative controls was presented for discussion.展开更多
The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four...The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.展开更多
文摘The control and prevention of pasteurellosis in rabbits which makes the hosts unsuitable for experimental use raised the needs to improve and simplify the procedures of Enzyme-Linked Immunosorbent Assay (ELISA) for detect of antibody against P.multocida. A comparison on the sensitivity and specificity of bacterial culture of antemortem and postmortem samples, complement fixation test and enzyme-linked immunosorbent assay of 11 apparently healthy adult rabbits was conducted. The incidence rates showed 45.45%,54.54% and 72.73% respectively. The sensitivity for the three methods were 0.63, 0.67,and 1.00,and specificity for them were 1.00, 0.67 and 1.00 respectively. Somatic serotypes of isolates of P.multocida from rabbits of three groups (rabbits of group 2 were with clinic signs, those of groups 1 and 3 were apparently healthy) revealed no remarkable differences,and the predominant types were type 3 and type 3, 4. This was somewhat different from the reports derived from other states. As the antigen of different serotype used in ELISA may have different sensitivity and specificity, which is affected also by different preparation method, a type non-specific antigen should be selected to meet such request. The trial of accomplishment of ELISA without positive and negative controls was presented for discussion.
基金Supported by the National Natural Science Foundation of China(31802345)China Postdoctoral Science Foundation(2018M630893)Heilongjiang Province Postdoctoral Science Foundation(LBH-Z18275)。
文摘The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.
