資源描述:
《記憶缺失性貝毒免疫膠體金快速檢測(cè)試紙條的研制》由會(huì)員上傳分享,免費(fèi)在線閱讀,更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、上海交通大學(xué)碩士學(xué)位論文分別為IgG1和IgG2a,通過體內(nèi)誘生腹水法獲得純化后效價(jià)為1:64000的抗軟骨藻酸單克隆抗體。3.膠體金及軟骨藻酸金標(biāo)抗體的制備采用檸檬酸三鈉還原氯金酸法,成功確立了20nm膠體金的制備條件,并利用制備好的膠體金,對(duì)前述制備的抗軟骨藻酸單克隆抗體進(jìn)行了膠體金標(biāo)記條件的摸索,成功確立了單克隆抗體的膠體金標(biāo)記條件:最適pH是9.0,最適標(biāo)記穩(wěn)定量是9.72μg/mL膠體金,并制備金標(biāo)抗體。4.軟骨藻酸膠體金免疫層析試紙條的制備對(duì)軟骨藻酸膠體金免疫層析試紙條的各項(xiàng)指標(biāo)進(jìn)行優(yōu)化,并確立了最佳條件。最終選擇SB06玻璃纖維素膜涂布4×稀釋度的金標(biāo)抗體構(gòu)成膠體金結(jié)合墊;選
2、擇Millipore180NC膜作為層析膜,羊抗鼠IgG(1mg/mL)和1μL包被抗原DA-BSA(0.867mg/mL)噴涂在NC膜上,分別作為質(zhì)控線和檢測(cè)線;選用SB08玻璃纖維素膜作為樣品墊;SX18吸水紙作為吸收墊;將膠體金結(jié)合墊、NC膜、樣品墊和吸收墊組裝成軟骨藻酸膠體金免疫層析試紙條5.軟骨藻酸膠體金免疫層析試紙條的應(yīng)用對(duì)膠體金免疫層析試紙條的檢測(cè)靈敏度、特異性和穩(wěn)定性進(jìn)行測(cè)試,結(jié)果表明,試紙條檢測(cè)時(shí)間不超過15min,檢測(cè)靈敏度達(dá)20ng/mL(對(duì)于貝類食品相當(dāng)于8μg/g貝肉),可以滿足國(guó)際規(guī)定的20μg/g的安全限值。特異性強(qiáng)、重復(fù)性好,4℃下可保存6個(gè)月。該方法靈敏度
3、高、簡(jiǎn)單快速,無需特殊儀器設(shè)備,適用于廣大基層單位的現(xiàn)場(chǎng)和批量檢測(cè),有著良好的商業(yè)開發(fā)應(yīng)用前景。關(guān)鍵詞:記憶缺失性貝毒,軟骨藻酸,單克隆抗體,膠體金,免疫層析第II頁(yè)萬方數(shù)據(jù)上海交通大學(xué)碩士學(xué)位論文DEVELOPMENTOFCOLLOIDALGOLDIMMUNOCHROMATOGRAPHICSTRIPFORRAPIDDETECTIONOFAMNESICSHELLFISHPOISONINGTOXINABSTRACTAmnesicShellfishPoisoning(ASP)isakindoftoxinthatproducedbytoxicredtidediatoms.Themaincompo
4、undinvolvedinthistoxicityisdomoicacid(DA),anexcitatoryneurotransmitteraminoacidofthekainicgroupthathadbeenisolatedfromamarinesponge.Domoicacidcanbebio-concentratedbyshellfishthroughthetransmissionoffoodchain,andcanalsobedischargedintomarinewaterdirectlybydiatoms.Peopleandothermarinemammalswhotakei
5、nthecontaminatedseafoodscouldsufferfromcentralnervoussystemdysfunction,notablylossofshort-termmemory,evencomaanddeath.Colloidalgoldimmunochromatographicassay(GICA)isanewimmunochromatographictechniqueinwhichacellulosemembraneisusedasthecarrier,andacolloidalgold-labeledantigenorantibodyisusedasthetr
6、acer.Itissimple,rapidandcheap,andtheresultisclearandeasytoanalyzewithoutanyequipment/facilitiesandskilledtechnicians.Sinceitisverysuitableforspot,rapiddetectionandbatchdetectiononsite,theGICAtechniquehasbecomeoneofthefastestgrowingimmunologicaldetectionmethodsandiswidelyusedforrapiddetectioninmari
7、netoxinsanalysis.Inthisstudy,thecolloidalgoldimmunochromatographicstripforrapiddetectionofdomoicacid(DA)wasdevelopedbyusingtheprincipleofmonoclonalantibodyandimmunochromatographicassay.Theresultswere第III頁(yè)萬方數(shù)據(jù)上海交通