玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)檢(jian)(jian)測(ce)(ce)(ce)試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he) 使(shi)(shi)用(yong)說(shuo)明書 （酶聯免(mian)(mian)疫法） 1 原(yuan)理(li)及(ji)(ji)(ji)用(yong)途(tu) 本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)采用(yong)間(jian)接(jie)競爭 ELISA 方(fang)法檢(jian)(jian)測(ce)(ce)(ce)谷物(wu)(wu)、飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)、食用(yong) 油(you)、啤酒、醬油(you)等樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)（Zearalenone，ZEN） （又(you)稱 F-2 毒素(su)(su)），試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)由預包被(bei)偶聯抗(kang)(kang)原(yuan)的(de)(de)(de)(de)(de)(de)(de)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)、辣根 酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)記物(wu)(wu)、抗(kang)(kang)體(ti)(ti)、標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)及(ji)(ji)(ji)其(qi)他(ta)配(pei)(pei)套試(shi)(shi)(shi)(shi)劑(ji)組(zu)成。檢(jian)(jian)測(ce)(ce)(ce)時(shi)(shi)，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)或(huo)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)溶(rong)液(ye)(ye)(ye)(ye)，樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)和(he)(he)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)上(shang)(shang)預包被(bei) 偶聯抗(kang)(kang)原(yuan)競爭抗(kang)(kang)玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)抗(kang)(kang)體(ti)(ti)，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)記物(wu)(wu)后(hou)，用(yong) TMB 底物(wu)(wu)顯色(se)，樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi)與其(qi)所(suo)含(han)玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)含(han)量(liang)成負(fu)相 關，與標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲(qu)(qu)線(xian)比較即(ji)(ji)可(ke)得出樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)(zhong)玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)的(de)(de)(de)(de)(de)(de)(de)殘(can)留量(liang)。 2 技術指標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao) 2.1 試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)靈敏度(du)(du)(du)(du)：0.3ppb(ng/ml) 2.2 反(fan)(fan)應(ying)(ying)(ying)(ying)模式：25℃，30min～15min 2.3 檢(jian)(jian)測(ce)(ce)(ce)下(xia)(xia)限： 谷物(wu)(wu)及(ji)(ji)(ji)其(qi)食品(pin)(pin)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao)…………………6ppb 飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)及(ji)(ji)(ji)飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao)………………………18ppb 玉(yu)(yu)米(mi)(mi)(mi)皮(pi)、麥(mai)麩…………………………18ppb 2.4 交(jiao)叉反(fan)(fan)應(ying)(ying)(ying)(ying)率(lv)(lv)： 玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)烯(xi)酮(tong)……………………100% 玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)酮(tong)………………………13% 玉(yu)(yu)米(mi)(mi)(mi)赤(chi)(chi)霉(mei)(mei)(mei)醇(chun)(chun)………………………＜1% 2.5 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)回收率(lv)(lv)： 谷物(wu)(wu)及(ji)(ji)(ji)其(qi)食品(pin)(pin)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao)…………………90%±15% 飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)及(ji)(ji)(ji)飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao)………………………80%±15% 玉(yu)(yu)米(mi)(mi)(mi)皮(pi)、麥(mai)麩…………………………80%±15% 3 試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)組(zu)成 酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)板(ban)(ban)(ban)……………………………96 孔(kong)(kong)(kong)(kong)(kong)(kong) 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)（黑蓋(gai)）：各 1ml 0ppb、0.3ppb、0.9ppb、2.7ppb、8.1ppb、24.3ppb 酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)記物(wu)(wu)（紅蓋(gai)）…………………5.5ml 抗(kang)(kang)體(ti)(ti)工(gong)作(zuo)(zuo)液(ye)(ye)(ye)(ye)（藍蓋(gai)）………………5.5ml 底物(wu)(wu)液(ye)(ye)(ye)(ye) A（白蓋(gai)）…………………6ml 底物(wu)(wu)液(ye)(ye)(ye)(ye) B（黑蓋(gai)）……………………6ml 終(zhong)(zhong)(zhong)止(zhi)(zhi)(zhi)(zhi)液(ye)(ye)(ye)(ye)（黃蓋(gai)）………………………6ml 20×濃(nong)縮(suo)洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)（白蓋(gai)）……………40ml 說(shuo)明書…………………………………1 份 蓋(gai)板(ban)(ban)(ban)膜…………………………………1 張 自封(feng)(feng)(feng)袋…………………………………1 個 4 需要(yao)的(de)(de)(de)(de)(de)(de)(de)器(qi)材(cai)和(he)(he)試(shi)(shi)(shi)(shi)劑(ji) 4.1 儀(yi)器(qi)：酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)、打印機、均(jun)質器(qi) 、振(zhen)蕩(dang)器(qi)、離(li)(li)(li)心(xin)機、刻 度(du)(du)(du)(du)移液(ye)(ye)(ye)(ye)管(guan)、天平(ping)(ping)（感量(liang) 0.01g） 4.2 微(wei)(wei)量(liang)移液(ye)(ye)(ye)(ye)器(qi)：單道(dao) 20µl-200µl，100µl-1000µl、多道(dao) 300µl 4.3 試(shi)(shi)(shi)(shi)劑(ji)：甲醇(chun)(chun)、去(qu)離(li)(li)(li)子(zi)水(shui)(shui) 5 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)前處理(li) 5.1 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)處理(li)前須知： 實驗(yan)器(qi)具必須潔(jie)凈(jing)并(bing)使(shi)(shi)用(yong)一(yi)(yi)次(ci)性(xing)吸(xi)(xi)(xi)(xi)頭(tou)，以避免(mian)(mian)污(wu)染干(gan)(gan)(gan)擾實 驗(yan)結(jie)果。 5.2 配(pei)(pei)液(ye)(ye)(ye)(ye)： 配(pei)(pei)液(ye)(ye)(ye)(ye) 1：樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)提取(qu)(qu)(qu)(qu)(qu)(qu)(qu)液(ye)(ye)(ye)(ye) 90%甲醇(chun)(chun)，即(ji)(ji) V 甲醇(chun)(chun):V 去(qu)離(li)(li)(li)子(zi)水(shui)(shui)=9：1。 配(pei)(pei)液(ye)(ye)(ye)(ye) 2：工(gong)作(zuo)(zuo)洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye) 將濃(nong)縮(suo)洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye) 20 倍稀釋(shi)(shi)(1 份洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)加(jia)(jia)(jia)(jia) 19 份去(qu)離(li)(li)(li)子(zi)水(shui)(shui))。 5.3 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)前處理(li)步驟： 5.3.1 谷物(wu)(wu)（大米(mi)(mi)(mi)、玉(yu)(yu)米(mi)(mi)(mi)、小米(mi)(mi)(mi)等低(di)(di)脂含(han)量(liang)）及(ji)(ji)(ji)其(qi)食品(pin)(pin)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao) 1）稱取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 2g 粉碎(sui)(sui)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)品(pin)(pin)于(yu)(yu)(yu)(yu) 50ml 離(li)(li)(li)心(xin)管(guan)中(zhong)(zhong)(zhong)，加(jia)(jia)(jia)(jia) 8ml 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)提取(qu)(qu)(qu)(qu)(qu)(qu)(qu)液(ye)(ye)(ye)(ye)， 振(zhen)蕩(dang) 5 分(fen)(fen)鐘(zhong)(zhong)(zhong)，室溫 4000 轉/分(fen)(fen)離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)(zhong)(zhong)； 2）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 0.5ml 上(shang)(shang)清，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 2ml 去(qu)離(li)(li)(li)子(zi)水(shui)(shui)，混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)； 3）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 50μl 進行(xing)(xing)分(fen)(fen)析。 稀釋(shi)(shi)倍數(shu)：20 檢(jian)(jian)測(ce)(ce)(ce)下(xia)(xia)限：6ppb 5.3.2 飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)及(ji)(ji)(ji)飼(si)料(liao)(liao)(liao)(liao)(liao)(liao)原(yuan)料(liao)(liao)(liao)(liao)(liao)(liao) 1）稱取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 2g 粉碎(sui)(sui)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)品(pin)(pin)于(yu)(yu)(yu)(yu) 50ml 離(li)(li)(li)心(xin)管(guan)中(zhong)(zhong)(zhong)，加(jia)(jia)(jia)(jia) 8ml 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)提取(qu)(qu)(qu)(qu)(qu)(qu)(qu)液(ye)(ye)(ye)(ye)， 振(zhen)蕩(dang) 5 分(fen)(fen)鐘(zhong)(zhong)(zhong)，室溫 4000 轉/分(fen)(fen)離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)(zhong)(zhong)； 2）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 0.5ml 上(shang)(shang)清，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 1ml 提取(qu)(qu)(qu)(qu)(qu)(qu)(qu)液(ye)(ye)(ye)(ye)，震蕩(dang)混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)； 3）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 0.5ml 混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)液(ye)(ye)(ye)(ye)體(ti)(ti)，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 2ml 去(qu)離(li)(li)(li)子(zi)水(shui)(shui)，混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)； 4）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 50μl 進行(xing)(xing)分(fen)(fen)析。 稀釋(shi)(shi)倍數(shu)：60 檢(jian)(jian)測(ce)(ce)(ce)下(xia)(xia)限：18ppb 5.3.3 玉(yu)(yu)米(mi)(mi)(mi)皮(pi)、麥(mai)麩等吸(xi)(xi)(xi)(xi)水(shui)(shui)性(xing)強的(de)(de)(de)(de)(de)(de)(de)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben) 1）稱取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 2g 粉碎(sui)(sui)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)品(pin)(pin)于(yu)(yu)(yu)(yu) 50ml 離(li)(li)(li)心(xin)管(guan)中(zhong)(zhong)(zhong)，加(jia)(jia)(jia)(jia) 24ml 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)提取(qu)(qu)(qu)(qu)(qu)(qu)(qu)液(ye)(ye)(ye)(ye)， 振(zhen)蕩(dang) 5 分(fen)(fen)鐘(zhong)(zhong)(zhong)，室溫 4000 轉/分(fen)(fen)離(li)(li)(li)心(xin) 10 分(fen)(fen)鐘(zhong)(zhong)(zhong)； 2）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 0.5ml 上(shang)(shang)清，加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 2ml 去(qu)離(li)(li)(li)子(zi)水(shui)(shui)，混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)； 3）取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 50μl 進行(xing)(xing)分(fen)(fen)析。 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)稀釋(shi)(shi)倍數(shu)：60 檢(jian)(jian)測(ce)(ce)(ce)下(xia)(xia)限：18ppb 注：由于(yu)(yu)(yu)(yu)毒素(su)(su)在(zai)(zai)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)分(fen)(fen)布呈非(fei)均(jun)勻(yun)(yun)狀(zhuang)態，取(qu)(qu)(qu)(qu)(qu)(qu)(qu)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)時(shi)(shi)需多點取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)充分(fen)(fen)混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)后(hou)再(zai)(zai)取(qu)(qu)(qu)(qu)(qu)(qu)(qu) 2g 進行(xing)(xing)試(shi)(shi)(shi)(shi)驗(yan)。 6 酶聯免(mian)(mian)疫試(shi)(shi)(shi)(shi)驗(yan)步驟 將所(suo)需試(shi)(shi)(shi)(shi)劑(ji)從 4℃冷藏環境中(zhong)(zhong)(zhong)取(qu)(qu)(qu)(qu)(qu)(qu)(qu)出，置(zhi)(zhi)于(yu)(yu)(yu)(yu)室溫平(ping)(ping)衡 30min 以上(shang)(shang), 洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)冷藏時(shi)(shi)可(ke)能(neng)會(hui)(hui)有(you)(you)(you)(you)結(jie)晶(jing)需恢復(fu)到(dao)室溫以充分(fen)(fen)溶(rong)解(jie)， 每(mei)(mei)種液(ye)(ye)(ye)(ye)體(ti)(ti)試(shi)(shi)(shi)(shi)劑(ji)使(shi)(shi)用(yong)前均(jun)須搖勻(yun)(yun)。取(qu)(qu)(qu)(qu)(qu)(qu)(qu)出需要(yao)數(shu)量(liang)的(de)(de)(de)(de)(de)(de)(de)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)板(ban)(ban)(ban)及(ji)(ji)(ji)框 架，將不用(yong)的(de)(de)(de)(de)(de)(de)(de)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)板(ban)(ban)(ban)放入(ru)(ru)(ru)自封(feng)(feng)(feng)袋，保存(cun)于(yu)(yu)(yu)(yu) 2-8℃。 6.1 編 號(hao)：將樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)和(he)(he)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)對應(ying)(ying)(ying)(ying)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)按序編號(hao)，每(mei)(mei)個樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben) 和(he)(he)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)做 2 孔(kong)(kong)(kong)(kong)(kong)(kong)平(ping)(ping)行(xing)(xing)，并(bing)記錄標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)孔(kong)(kong)(kong)(kong)(kong)(kong)和(he)(he)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)孔(kong)(kong)(kong)(kong)(kong)(kong)所(suo)在(zai)(zai)的(de)(de)(de)(de)(de)(de)(de)位(wei)置(zhi)(zhi)。 6.2 加(jia)(jia)(jia)(jia)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)反(fan)(fan)應(ying)(ying)(ying)(ying)：加(jia)(jia)(jia)(jia)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)品(pin)(pin)或(huo)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben) 50µl/孔(kong)(kong)(kong)(kong)(kong)(kong)到(dao)各自的(de)(de)(de)(de)(de)(de)(de)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)中(zhong)(zhong)(zhong)，然(ran) 后(hou)加(jia)(jia)(jia)(jia)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)記物(wu)(wu) 50µl/孔(kong)(kong)(kong)(kong)(kong)(kong)，再(zai)(zai)加(jia)(jia)(jia)(jia)入(ru)(ru)(ru) 50µl/孔(kong)(kong)(kong)(kong)(kong)(kong)的(de)(de)(de)(de)(de)(de)(de)抗(kang)(kang)體(ti)(ti)工(gong)作(zuo)(zuo)液(ye)(ye)(ye)(ye)，用(yong)蓋(gai) 板(ban)(ban)(ban)膜封(feng)(feng)(feng)板(ban)(ban)(ban)，輕輕振(zhen)蕩(dang) 5 秒(miao)(miao)混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)，25℃反(fan)(fan)應(ying)(ying)(ying)(ying) 30 分(fen)(fen)鐘(zhong)(zhong)(zhong)。 6.3 洗(xi)(xi) 滌(di)：小心(xin)揭開蓋(gai)板(ban)(ban)(ban)膜，甩去(qu)孔(kong)(kong)(kong)(kong)(kong)(kong)內(nei)液(ye)(ye)(ye)(ye)體(ti)(ti)，每(mei)(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia) 350ul 工(gong)作(zuo)(zuo)洗(xi)(xi)滌(di)液(ye)(ye)(ye)(ye)，靜置(zhi)(zhi) 30 秒(miao)(miao)后(hou)棄去(qu)，重(zhong)復(fu)洗(xi)(xi)滌(di) 5 次(ci)，最(zui)后(hou)一(yi)(yi)次(ci)拍 干(gan)(gan)(gan)（用(yong)吸(xi)(xi)(xi)(xi)水(shui)(shui)紙拍干(gan)(gan)(gan)，拍干(gan)(gan)(gan)后(hou)未被(bei)清除的(de)(de)(de)(de)(de)(de)(de)氣泡可(ke)用(yong)干(gan)(gan)(gan)凈(jing)的(de)(de)(de)(de)(de)(de)(de)槍頭(tou)刺 破(po)）。 6.4 顯 色(se)：每(mei)(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)入(ru)(ru)(ru)底物(wu)(wu)液(ye)(ye)(ye)(ye) A 50µl，再(zai)(zai)加(jia)(jia)(jia)(jia)底物(wu)(wu)液(ye)(ye)(ye)(ye) B 50µl， 輕輕振(zhen)蕩(dang) 5 秒(miao)(miao)混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)，25℃避光(guang)(guang)(guang)顯色(se) 15 分(fen)(fen)鐘(zhong)(zhong)(zhong)（若藍色(se)過(guo)(guo)淺(qian)，可(ke) 適當延(yan)長反(fan)(fan)應(ying)(ying)(ying)(ying)時(shi)(shi)間(jian)）。產(chan)(chan)(chan)品(pin)(pin)縮(suo)寫：ZEN 貨號(hao)：RJ-P100055 版本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)：2022.1 2 6.5 終(zhong)(zhong)(zhong) 止(zhi)(zhi)(zhi)(zhi)：每(mei)(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)加(jia)(jia)(jia)(jia)入(ru)(ru)(ru)終(zhong)(zhong)(zhong)止(zhi)(zhi)(zhi)(zhi)液(ye)(ye)(ye)(ye) 50µl，輕輕振(zhen)蕩(dang)混(hun)(hun)(hun)(hun)(hun)(hun)勻(yun)(yun)，終(zhong)(zhong)(zhong)止(zhi)(zhi)(zhi)(zhi)反(fan)(fan) 應(ying)(ying)(ying)(ying)。6.6 測(ce)(ce)(ce)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)值(zhi)(zhi)(zhi)：用(yong)酶標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)儀(yi)于(yu)(yu)(yu)(yu) 450nm 處測(ce)(ce)(ce)定每(mei)(mei)孔(kong)(kong)(kong)(kong)(kong)(kong)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi)（建(jian)議 用(yong)雙(shuang)波長 450/630nm）。測(ce)(ce)(ce)定應(ying)(ying)(ying)(ying)在(zai)(zai)終(zhong)(zhong)(zhong)止(zhi)(zhi)(zhi)(zhi)反(fan)(fan)應(ying)(ying)(ying)(ying)后(hou) 10 分(fen)(fen)鐘(zhong)(zhong)(zhong)內(nei)完成。 7 結(jie)果分(fen)(fen)析 7.1 百分(fen)(fen)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)率(lv)(lv)的(de)(de)(de)(de)(de)(de)(de)計(ji)(ji)算 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)或(huo)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)(de)百分(fen)(fen)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)率(lv)(lv)等于(yu)(yu)(yu)(yu)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)或(huo)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)(de)百分(fen)(fen)吸(xi)(xi)(xi)(xi) 光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi)的(de)(de)(de)(de)(de)(de)(de)平(ping)(ping)均(jun)值(zhi)(zhi)(zhi)（雙(shuang)孔(kong)(kong)(kong)(kong)(kong)(kong)）除以第一(yi)(yi)個標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)（0ppb）的(de)(de)(de)(de)(de)(de)(de)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du) 值(zhi)(zhi)(zhi)，再(zai)(zai)乘(cheng)以 100%，即(ji)(ji) 百分(fen)(fen)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi)（%）＝ A ×100% A0 A—標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)溶(rong)液(ye)(ye)(ye)(ye)或(huo)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)溶(rong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)(de)平(ping)(ping)均(jun)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi) A0—0ppb 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)溶(rong)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)(de)平(ping)(ping)均(jun)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)度(du)(du)(du)(du)值(zhi)(zhi)(zhi) 7.2 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲(qu)(qu)線(xian)的(de)(de)(de)(de)(de)(de)(de)繪制與計(ji)(ji)算 以標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)百分(fen)(fen)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)率(lv)(lv)為(wei)縱坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)，對應(ying)(ying)(ying)(ying)的(de)(de)(de)(de)(de)(de)(de)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)濃(nong)度(du)(du)(du)(du)（ppb） 的(de)(de)(de)(de)(de)(de)(de)對數(shu)為(wei)橫坐(zuo)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)，繪制標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)液(ye)(ye)(ye)(ye)的(de)(de)(de)(de)(de)(de)(de)半對數(shu)曲(qu)(qu)線(xian)圖(tu)。將樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)(de)百分(fen)(fen) 吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang)率(lv)(lv)代入(ru)(ru)(ru)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲(qu)(qu)線(xian)中(zhong)(zhong)(zhong)，從標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲(qu)(qu)線(xian)上(shang)(shang)讀出樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)所(suo)對應(ying)(ying)(ying)(ying)的(de)(de)(de)(de)(de)(de)(de)濃(nong) 度(du)(du)(du)(du)，乘(cheng)以其(qi)對應(ying)(ying)(ying)(ying)的(de)(de)(de)(de)(de)(de)(de)稀釋(shi)(shi)倍數(shu)即(ji)(ji)為(wei)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)中(zhong)(zhong)(zhong)待測(ce)(ce)(ce)物(wu)(wu)的(de)(de)(de)(de)(de)(de)(de)實際(ji)濃(nong)度(du)(du)(du)(du)。 若利用(yong)試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)專(zhuan)業分(fen)(fen)析軟件進行(xing)(xing)計(ji)(ji)算，更便于(yu)(yu)(yu)(yu)大量(liang)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)的(de)(de)(de)(de)(de)(de)(de) 準(zhun)(zhun)(zhun)(zhun)(zhun)確(que)、快速分(fen)(fen)析。（歡迎來電索取(qu)(qu)(qu)(qu)(qu)(qu)(qu)） 8 注意事項 8.1 室溫低(di)(di)于(yu)(yu)(yu)(yu) 25℃或(huo)試(shi)(shi)(shi)(shi)劑(ji)及(ji)(ji)(ji)樣(yang)(yang)(yang)(yang)(yang)(yang)(yang)本(ben)(ben)(ben)(ben)(ben)(ben)(ben)(ben)沒有(you)(you)(you)(you)回到(dao)室溫（25℃）會(hui)(hui)導致 所(suo)有(you)(you)(you)(you)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)的(de)(de)(de)(de)(de)(de)(de) OD 值(zhi)(zhi)(zhi)偏(pian)低(di)(di)。 8.2 在(zai)(zai)洗(xi)(xi)板(ban)(ban)(ban)過(guo)(guo)程中(zhong)(zhong)(zhong)如果出現(xian)(xian)板(ban)(ban)(ban)孔(kong)(kong)(kong)(kong)(kong)(kong)干(gan)(gan)(gan)燥的(de)(de)(de)(de)(de)(de)(de)情況(kuang)，則會(hui)(hui)出現(xian)(xian)標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)曲(qu)(qu) 線(xian)不成線(xian)性(xing)，重(zhong)復(fu)性(xing)不好的(de)(de)(de)(de)(de)(de)(de)現(xian)(xian)象。所(suo)以洗(xi)(xi)板(ban)(ban)(ban)拍干(gan)(gan)(gan)后(hou)應(ying)(ying)(ying)(ying)立即(ji)(ji)進行(xing)(xing) 下(xia)(xia)一(yi)(yi)步操作(zuo)(zuo)。 8.3 混(hun)(hun)(hun)(hun)(hun)(hun)合(he)要(yao)均(jun)勻(yun)(yun)，洗(xi)(xi)板(ban)(ban)(ban)要(yao)，在(zai)(zai) ELISA 分(fen)(fen)析中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)再(zai)(zai)現(xian)(xian)性(xing)，很(hen) 大程度(du)(du)(du)(du)上(shang)(shang)取(qu)(qu)(qu)(qu)(qu)(qu)(qu)決(jue)于(yu)(yu)(yu)(yu)洗(xi)(xi)板(ban)(ban)(ban)的(de)(de)(de)(de)(de)(de)(de)一(yi)(yi)致性(xing)。 8.4 在(zai)(zai)所(suo)有(you)(you)(you)(you)孵(fu)育過(guo)(guo)程中(zhong)(zhong)(zhong)，用(yong)蓋(gai)板(ban)(ban)(ban)膜封(feng)(feng)(feng)住(zhu)微(wei)(wei)孔(kong)(kong)(kong)(kong)(kong)(kong)板(ban)(ban)(ban)，避免(mian)(mian)光(guang)(guang)(guang)線(xian)照射。 8.5 不要(yao)使(shi)(shi)用(yong)過(guo)(guo)了有(you)(you)(you)(you)效(xiao)期的(de)(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)，不要(yao)交(jiao)換(huan)使(shi)(shi)用(yong)不同批號(hao)試(shi)(shi)(shi)(shi) 劑(ji)盒(he)(he)中(zhong)(zhong)(zhong)的(de)(de)(de)(de)(de)(de)(de)試(shi)(shi)(shi)(shi)劑(ji)。 8.6 顯色(se)液(ye)(ye)(ye)(ye)若有(you)(you)(you)(you)任何顏色(se)表(biao)明變質，應(ying)(ying)(ying)(ying)當棄之(zhi)。0 標(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)(biao)準(zhun)(zhun)(zhun)(zhun)(zhun)的(de)(de)(de)(de)(de)(de)(de)吸(xi)(xi)(xi)(xi)光(guang)(guang)(guang) 度(du)(du)(du)(du)值(zhi)(zhi)(zhi)小于(yu)(yu)(yu)(yu) 0.5 個單位(wei)（A450nm< 0.5 ）時(shi)(shi)，表(biao)示試(shi)(shi)(shi)(shi)劑(ji)可(ke)能(neng)變質。 8.7 反(fan)(fan)應(ying)(ying)(ying)(ying)終(zhong)(zhong)(zhong)止(zhi)(zhi)(zhi)(zhi)液(ye)(ye)(ye)(ye)有(you)(you)(you)(you)腐(fu)蝕性(xing)，避免(mian)(mian)接(jie)觸皮(pi)膚。 9 貯藏及(ji)(ji)(ji)保存(cun)期 儲藏條件：試(shi)(shi)(shi)(shi)劑(ji)盒(he)(he)于(yu)(yu)(yu)(yu) 2-8℃保存(cun)，避免(mian)(mian)冷凍。 保 質 期：該產(chan)(chan)(chan)品(pin)(pin)有(you)(you)(you)(you)效(xiao)期為(wei) 1 年，生產(chan)(chan)(chan)日期見包裝(zhuang)盒(he)(he)。