畜禽養殖場作為動物疫病傳播的源頭,在疫病發生時及時進行病原診斷,采取相應的應對措施,是目前動物疫病防控的關鍵環節。
muqianguoneidongwuyibingdebingyuanxuezhenduanshijixuanzefanweiyouxian。erzaitongyongdexueqingxuejianceshijizhong,juedaduoshushijiancexueqingzhongkangtidemianyishiji,qijieguogengduofanyingleyimiaomianyixiaoguo,duibingyuanganranzhinengjianjiefenxi,wufaquezhen,hekuangdongwuzaizaoshouganranhou,bingduyaozaitineiqianfuyiduanshijian,dongwucaihuichanshengkangti,jiancejieguowangwangzhihoubingyuanganran。nengduiyeduganranjinxingjianbiedechengshukangtijianceshijizhiyouzhuweikuangquandegE抗kang體ti試shi劑ji盒he,由you於yu疫yi苗miao的de純chun化hua工gong藝yi問wen題ti,口kou蹄ti疫yi的de非fei結jie構gou蛋dan白bai的de抗kang體ti目mu前qian無wu法fa產chan生sheng理li論lun上shang的de鑒jian別bie診zhen斷duan作zuo用yong。另ling一yi方fang麵mian能neng監jian測ce疫yi病bing病bing原yuan的de試shi劑ji的de商shang品pin化hua試shi劑ji偏pian少shao。免mian疫yi熒ying光guang抗kang體ti技ji術shu受shou限xian於yu基ji層ceng養yang殖zhi場chang熒ying光guang顯xian微wei鏡jing以yi及ji熒ying光guang標biao記ji抗kang體ti的de缺que乏fa,故gu在zai多duo數shu基ji層ceng養yang殖zhi試shi驗yan室shi難nan以yi普pu及ji。由you於yu達da到dao市shi場chang需xu求qiu的de特te異yi性xing及ji靈ling敏min度du並bing能neng配pei對dui的de動dong物wu疫yi病bing抗kang體ti的de缺que乏fa,免mian疫yi試shi劑ji中zhong目mu前qian的de病bing原yuan檢jian測ce產chan品pin很hen少shao,這zhe類lei產chan品pin開kai發fa的de周zhou期qi長chang,不bu確que定ding性xing很hen大da,很hen難nan應ying對dui基ji層ceng疫yi病bing防fang控kong的de需xu要yao。因yin此ci養yang殖zhi場chang試shi驗yan室shi亟ji需xu合he適shi的de病bing原yuan學xue檢jian測ce試shi劑ji。
鑒於基於核酸擴增的分子診斷技術發展迅速,熒光PCR及電泳PCR已經被農業部及各省市疫控單位逐步采用,扭轉了疫病診斷中病原診斷技術空白的態勢。但對養殖場基層試驗室來說,PCR分子診斷技術的使用存在不少阻礙:
1、分子診斷試劑一般要冷凍保存,成本高且運輸半徑有限;
2、核酸提取步驟複雜緩慢,基層技術人員操作困難;
3、基層單位無力購買價格偏高的PCR擴增儀,基層技術人員也不容易掌握儀器操作要領。因為這些難點的存在,分子診斷技術一直沒有在我國養殖基層試驗室普及。
1、分子診斷試劑一般要冷凍保存,成本高且運輸半徑有限;
2、核酸提取步驟複雜緩慢,基層技術人員操作困難;
3、基層單位無力購買價格偏高的PCR擴增儀,基層技術人員也不容易掌握儀器操作要領。因為這些難點的存在,分子診斷技術一直沒有在我國養殖基層試驗室普及。
快靈生物開發的基因試紙卡,開啟了現場分子診斷步入養殖場的新時代。它包括基因提取試劑、恒溫擴增試劑及免疫層析試紙卡三部分,能常溫運輸;在常溫下同時完成 DNA/RNA樣品提取,僅需微型離心機或專利磁力架10分鍾內完成操作;添加樣品模板後將反應管密閉後放置在變溫金屬浴上保溫60分鍾,擴增完成後將反應管插入試紙卡槽後一步按壓就啟動溶液層析,3-5fenzhonghoujikemushicengxishizhidexiantiaoxiansepandingjieguo。zhuanlishenqingzhongdejiyinshizhikajiejuelekuozengchanwurongyejiqiqirongjiaodexielouwurandaozhihouxuyangpinjiaochawurandewenti,kaiqilexianchangfenzizhenduanburulejicengyangzhichangdexinshidai。jiyinshizhikanengzhijiejiancedaobingyuan,henshiheduijiankangxiedupaidudedongwujinxingshaizha,yeshiheduiganranzaoqidedongwujinxingquezhen,dadasuoduanchuangkouqi。ciwai,duiyuhanjianbingyuanhuoturanbaofadexinbingyuan,zhiyaonengzaijiyinkuzhongzhaxundaogaibingyuandejiyinxulie,jiyinshizhikanengzuikuaizaiyizhouneiyanfachulai。gaijiyinshizhikadecaozuoguochengyizhuanlicilijia,bianwenjinshuyu,liangzhiyiyeqiangweilishuomingruxiatu。


快靈生物的基因試紙卡檢測盒的特點在於:
1.簡單快速提取核酸:快靈生物的核酸提取試劑在常溫下10分鍾內能同時完成樣品中DNA與RNA的提取。目前基因變異最快,危害性最大的疫病往往都是RNA病毒引起的。其他商業化RNA病bing毒du提ti取qu試shi劑ji一yi般ban要yao求qiu在zai低di溫wen冷leng凍dong條tiao件jian下xia提ti取qu操cao作zuo,基ji層ceng單dan位wei往wang往wang無wu法fa實shi現xian。快kuai靈ling生sheng物wu經jing過guo無wu數shu次ci的de實shi驗yan摸mo索suo,終zhong於yu配pei製zhi了le獨du特te的de基ji因yin提ti取qu試shi劑ji,僅jin需xu微wei型xing高gao速su離li心xin機ji或huo專zhuan利li磁ci力li架jia,10分鍾內就能完成單份樣品的核酸提取。我們已經在細菌病毒樣品,全血與組織樣品分別進行了DNA、RNA的提取實驗,結果都很良好。簡單的RNA提取試劑是基因試紙卡步入基層應用的基礎。
2. 擴增核酸的高靈敏高特異的快速免疫檢測技術:快kuai靈ling生sheng物wu通tong過guo膠jiao體ti金jin試shi紙zhi實shi現xian了le基ji因yin擴kuo增zeng產chan物wu的de高gao靈ling敏min高gao特te異yi檢jian測ce。膠jiao體ti金jin試shi紙zhi具ju有you極ji高gao的de的de檢jian測ce靈ling敏min度du,如ru克ke倫lun特te羅luo的de產chan品pin能neng達da到dao納na克ke級ji別bie(單位ng/ml),erjiyinkuozengjishuzekeyixunsujiangchaohenliangdebajiyinteyikuozengbaiwanbei。kuailingshengwujiangjiaotijinjishuyujiyinkuozengjishuwanmeijiehe,shixianlegaolingmindehesuanjiance。wogongsiduizhaoshiyanbiaoming,dangPCR擴增產物在瓊脂糖凝膠電泳上呈現微弱信號時,快靈生物的基因試紙卡對稀釋100倍的擴增產物仍有明顯信號。當LAMP恒溫擴增溶液采用染料(如SYBR Green I)隻能檢測到微弱信號時,快靈生物基因試紙卡對100倍(bei)稀(xi)釋(shi)後(hou)的(de)擴(kuo)增(zeng)溶(rong)液(ye)仍(reng)有(you)明(ming)顯(xian)信(xin)號(hao)。因(yin)此(ci)基(ji)因(yin)試(shi)紙(zhi)卡(ka)能(neng)實(shi)現(xian)更(geng)高(gao)靈(ling)敏(min)度(du)的(de)信(xin)號(hao)檢(jian)測(ce),或(huo)者(zhe)在(zai)同(tong)樣(yang)的(de)產(chan)物(wu)濃(nong)度(du)下(xia),能(neng)保(bao)證(zheng)更(geng)好(hao)的(de)信(xin)號(hao)判(pan)別(bie)。更(geng)重(zhong)要(yao)的(de)是(shi)染(ran)料(liao)檢(jian)測(ce)方(fang)法(fa)沒(mei)有(you)特(te)異(yi)性(xing),隻(zhi)要(yao)溶(rong)液(ye)中(zhong)存(cun)在(zai)一(yi)定(ding)濃(nong)度(du)的(de)核(he)酸(suan)就(jiu)有(you)信(xin)號(hao),無(wu)法(fa)區(qu)分(fen)信(xin)號(hao)是(shi)來(lai)自(zi)特(te)異(yi)性(xing)擴(kuo)增(zeng)產(chan)物(wu)還(hai)是(shi)非(fei)特(te)異(yi)性(xing)擴(kuo)增(zeng)產(chan)物(wu)。而(er)基(ji)因(yin)試(shi)紙(zhi)卡(ka)的(de)檢(jian)測(ce)則(ze)是(shi)引(yin)物(wu)與(yu)探(tan)針(zhen)雙(shuang)重(zhong)保(bao)證(zheng)擴(kuo)增(zeng)產(chan)物(wu)的(de)特(te)異(yi)性(xing),能(neng)很(hen)好(hao)地(di)區(qu)分(fen)非(fei)特(te)異(yi)擴(kuo)增(zeng)產(chan)物(wu)。
3. 精致的防汙染基因試紙卡:快靈生物專利申請中的防汙染基因試紙卡,是目前解決基層單位基因擴增產物汙染的最佳方案。盡管在熒光定量PCR和LAMP等deng方fang法fa中zhong,可ke以yi不bu開kai蓋gai檢jian測ce,但dan其qi使shi用yong的de反fan應ying管guan並bing沒mei有you嚴yan格ge密mi封feng的de結jie構gou設she計ji,滲shen漏lou與yu意yi外wai開kai蓋gai仍reng然ran可ke能neng導dao致zhi汙wu染ran。快kuai靈ling生sheng物wu專zhuan利li申shen請qing中zhong的de防fang汙wu染ran基ji因yin試shi紙zhi卡ka的de塑su料liao套tao件jian中zhong存cun在zai多duo級ji密mi封feng設she計ji,並bing能neng承cheng受shou一yi定ding的de空kong氣qi壓ya力li差cha,反fan應ying管guan內nei的de擴kuo增zeng產chan物wu被bei封feng存cun在zai試shi紙zhi卡ka中zhong,即ji使shi層ceng析xi試shi紙zhi上shang的de擴kuo增zeng產chan物wu形xing成cheng了le氣qi溶rong膠jiao也ye無wu法fa擴kuo散san出chu來lai。
4. 一步操作的核酸恒溫擴增技術:快kuai靈ling生sheng物wu采cai用yong了le一yi步bu操cao作zuo的de核he酸suan恒heng溫wen擴kuo增zeng技ji術shu,適shi合he基ji層ceng單dan位wei操cao作zuo人ren員yuan使shi用yong。近jin年nian開kai發fa出chu了le多duo種zhong恒heng溫wen擴kuo增zeng技ji術shu,有you些xie技ji術shu需xu要yao進jin行xing多duo步bu操cao作zuo才cai實shi現xian核he酸suan(DNA或RNA)的擴增。快靈生物公司綜合了LAMP技術與RCA技術的特點,獨創了新型的恒溫擴增技術,能一次添加相關試劑與樣品後,在50℃/64℃兩個溫度下共反應60分鍾就能達到層析試紙需要的信號濃度。
5. 基因試紙卡的常溫運輸:快kuai靈ling生sheng物wu的de基ji因yin試shi紙zhi卡ka檢jian測ce盒he中zhong的de試shi劑ji與yu物wu料liao都dou能neng在zai常chang溫wen下xia運yun輸shu,解jie決jue了le基ji因yin檢jian測ce試shi劑ji對dui基ji層ceng單dan位wei的de物wu流liu障zhang礙ai。目mu前qian絕jue大da部bu分fen的de基ji因yin檢jian測ce試shi劑ji都dou需xu要yao在zai-20℃detiaojianxiacaiyongganbingyunshu,shidechanpindepeisongheshiyongjiweibubian。zuoweinongmuyedejicengdanwei,chuyuyibingfangkongdexuyao,chuqinyangzhichangwangwangdichujiaoquhuopianyuandexiangcun。jiyinjianceshijidechangwenyunshu,shijiyinjiancejishujinruzhexiejicengdanweidexianjuetiaojian。kuailingchanpinzecaiyongduyoudebaohuji,duijianceshijizhongbuyibaocundechengfenjinxingdongganguhua,wanquanshixianlechanpindechangwenyunshu,geiguangdayangzhichangkaiqilejiyinjiancejishudedamen。
6. 最適應基層人員使用的方案:kuailingshengwudejiyinshizhikajiancehedequanliuchengcaozuojiandanfangbian,jishizhongxuewenhuarenyuanjingjiandanpeixunjiunengzhangwo。yinciduiguangdadejicengdanweikehu,caiyongjiyinshizhikajinxingyibingjiance,wanquanbuxuyaoewaizhaopinzhuanyedejishurenyuan。tongshijiyinshizhikasuoxupeitaoyiqizhiyouweixinggaosulixinjihuozhuanlicilijia,bianwenjinshuyuheliangzhiyiyeqiang,xuyaodepeizhifeiyongjiaodi,juedabufendejicengdanweidounengfudandeqi。
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