A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized...A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody,and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip,the capture antibody was laid on a sample pad,the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C,Swine vesicular disease (SVD),Vesicular stomatiti svirus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically,the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple,easy and fast for clinical testing on field sites; no special instruments and skills are required,and the result can be obtained within 15 min. To our knowledge,this is the first rapid immunochromatogarpic assay for serotype A of FMDV.展开更多
为研究副猪嗜血杆菌(HPS)的耐药性和耐药机制,本研究从临床疑似患多发浆膜炎和关节炎猪的病料样品中分离到30株致病菌,对其采用16S r RNA菌株鉴定方法和KRG琼脂扩散血清分型方法进行鉴定,并利用琼脂稀释法和PCR方法对分离株进行12种常...为研究副猪嗜血杆菌(HPS)的耐药性和耐药机制,本研究从临床疑似患多发浆膜炎和关节炎猪的病料样品中分离到30株致病菌,对其采用16S r RNA菌株鉴定方法和KRG琼脂扩散血清分型方法进行鉴定,并利用琼脂稀释法和PCR方法对分离株进行12种常用抗菌药物的最小抑菌浓度(MIC)测定和相应耐药基因或突变位点的检测。鉴定结果显示,30株分离株均为HPS,其血清型主要为5型(26.7%)、13型(26.7%)和不可分型(26.7%)。药物敏感性试验结果显示,30株HPS对氨苄西林、泰妙菌素、恩诺沙星和亚胺培南的耐药率较高,分别为50%、50%、46.7%和100%。利用PCR重点检测了介导喹诺酮类药物的耐药基因和基因突变情况,结果显示qnr A的检出率高达98.7%。根据MIC试验结果从中选出14株耐药性比较强的菌株,检测其gyr A/gyr B/par C/par E 4种基因的突变位点,结果显示gyr A基因突变位点主要为S83F和D87N,gyr B的突变位点均为P472G,par C的突变位点主要为L73S和A77E,par E基因的突变位点主要为R254H和A227T。本研究结果为临床选用合理抗菌药物防治HPS病提供了实验依据。展开更多
基金Financial supported by the Gansu ProvincialSci. & Tech. Department (1002NKDA037)
文摘A sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (FMDV) serotypes was developed. In this rapid test,affinity purified polyclonal antibodies from Guinea pigs which were immunized with sucking-mouse adapted FMD virus (A/AV88(L) strain) were conjugated to colloidal gold beads and used as the capture antibody,and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted FMD virus (A/CHA/09 strain) were used as detector antibody. On the nitrocellulose membrane of the immunochromatographic strip,the capture antibody was laid on a sample pad,the detector antibody was printed at the test line(T) and goat anti-guinea pigs IgG antibodies were immobilized to the control line(C). The lower detection limit of the test for a FMDV 146S antigen is 11.7ng/ml as determined in serial tests after the strip device was assembled and the assay condition optimization. No cross reactions were found with FMDV serotype C,Swine vesicular disease (SVD),Vesicular stomatiti svirus (VSV) and vesicular exanthema of swine virus (VES) viral antigens with this rapid test. Clinically,the diagnostic sensitivity of this test for FMDV serotypes A was 88.7% which is as same as an indirect-sandwich ELISA. The specificity of this strip test was 98.2% and is comparable to the 98.7% obtained with indirect-sandwich ELISA. This rapid strip test is simple,easy and fast for clinical testing on field sites; no special instruments and skills are required,and the result can be obtained within 15 min. To our knowledge,this is the first rapid immunochromatogarpic assay for serotype A of FMDV.
