Ukusetyenziswa kokuxilongwa kwi-vitro kuphela

Nceda ufunde le phakheji ngononophelo phambi kokuba uyisebenzise kwaye ulandele imiyalelo ngokungqongqo. Ukuthembeka kweziphumo zovavanyo akunakuqinisekiswa ukuba kukho naluphi na uphambuko kwimiyalelo ekwesi sithuba sephakheji.

UKUSETYENZISWA OKUJOLISWEYO

Ikhithi yokuxilonga ye-Hepatitis C Virus Antibody (Fluorescence Immunochromatographic Assay) luvavanyo lwe-fluorescence immunochromatographic lokubona ubungakanani be-HCV antibody kwi-serum okanye kwi-plasma yomntu, nto leyo ebaluleke kakhulu kwixabiso loxilongo losulelo lwe-hepatitis C. Zonke iisampulu ezilungileyo kufuneka ziqinisekiswe zezinye iindlela. Olu vavanyo lwenzelwe ukusetyenziswa ziingcali zempilo kuphela.

1. Beka ecaleni zonke ii-reagents kunye neesampuli ukuya kubushushu begumbi.
2.Vula i-Portable Immune Analyzer (WIZ-A101), faka igama lokugqitha le-akhawunti ngokwendlela yokusebenza kwesixhobo, uze ufake i-interface yokubona.
3.Skena ikhowudi yokuchonga ukuze uqinisekise into yovavanyo.
4.Khupha ikhadi lovavanyo kwingxowa yefoyile.
5.Faka ikhadi lovavanyo kwindawo yokubeka ikhadi, skena ikhowudi ye-QR, uze umisele into yovavanyo.
6. Yongeza isampuli yeserum okanye iplasma engama-20μL kwisampuli ye-diluent, uze udibanise kakuhle.
7. Yongeza isisombululo sesampulu se-80μL kwisampulu yomthombo wekhadi.
8.Cofa iqhosha elithi “uvavanyo oluqhelekileyo”, emva kwemizuzu eli-15, isixhobo siza kuzibona ngokuzenzekelayo ikhadi lovavanyo, singafunda iziphumo kwisikrini sokubonisa isixhobo, size sirekhode/siprinte iziphumo zovavanyo.
9. Jonga imiyalelo ye-Portable Immune Analyzer (WIZ-A101).

ISISHWANKATHELO

Intsholongwane yeHepatitis C (HCV) yintsholongwane ye-RNA (9.5 kb) ekwintsapho yeFlaviviridae. Ii-genotypes ezintandathu eziphambili kunye nothotho lwee-subtypes ze-HCV zichongiwe. I-HCV, eyahlulwe ngo-1989, ngoku iyaziwa njengesizathu esiphambili se-hepatitis engeyiyo i-A, engeyiyo i-B enxulumene notofelo-gazi. Esi sifo sibonakala ngesimo esibukhali nesingapheliyo. Ngaphezulu kwama-50% abantu abosulelekileyo bahlakulela i-hepatitis engapheliyo enzima, esongela ubomi kunye ne-cirrhosis yesibindi kunye ne-hepatocellular carcinomas. Ukusukela oko kwaqaliswa ngo-1990 ukuhlolwa kwegazi okuchasene ne-HCV, ukwanda kolu sulelo kubantu abafumana utofelo-gazi kuye kwehla kakhulu. Izifundo zeklinikhi zibonisa ukuba inani elikhulu labantu abosulelekileyo yi-HCV livelisa ii-antibodies kwi-NS5 non-structural protein yentsholongwane. Ngenxa yoku, uvavanyo luquka ii-antigens ezivela kummandla we-NS5 we-genome yentsholongwane ukongeza kwi-NS3 (c200), NS4 (c200) kunye neCore (c22).

UMGAQO WENKQUBO

I-membrane yesixhobo sovavanyo igqunywe nge-antigen ye-HCV kwindawo yovavanyo kunye ne-antibody ye-IgG yebhokhwe kwindawo yokulawula. Iileyibhile zigqunywe nge-antigen ye-HCV ebhalwe nge-fluorescence kunye ne-IgG yerabbit kwangaphambili. Xa kuvavanywa isampuli elungileyo, i-antibody ye-HCV kwisampuli idibana ne-antigen ye-HCV ebhalwe nge-fluorescence, kwaye yenze umxube wokuzikhusela. Phantsi kwesenzo se-immunochromatography, ukuhamba okuntsonkothileyo kwicala lephepha elifunxayo, xa i-complex idlula kwindawo yovavanyo, idibene ne-antigen ye-HCV yokugquma i-antigen, yenza i-complex entsha. Inqanaba le-antibody ye-HCV lihambelana kakuhle nesignali ye-fluorescence, kwaye uxinzelelo lwe-antibody ye-HCV kwisampuli lunokubonwa ngovavanyo lwe-immunoassay ye-fluorescence.

IZINTO EZINIKEZELWAYO NEZIXHOBO

Izixhobo zephakheji ze-25T：
Ikhadi lovavanyo ngalinye lifakwe ifoyile ngesixhobo sokucoca
.Iisampulu zokuxuba
.Iphakheji efakiweyo

IZIXHOBO EZIFUNEKAYO KODWA AZINIKELWANGA
Isikhongozeli sokuqokelela isampuli, isibali-xesha
IMFUNDO YOKUQOKELELA NOKUGCINA IMIZEKELO
1.Iisampulu ezivavanyiweyo zinokuba yi-serum, i-heparin anticoagulant plasma okanye i-EDTA anticoagulant plasma.
2. Ngokweendlela eziqhelekileyo zokuqokelela isampuli. Isampuli ye-serum okanye yeplasma ingagcinwa efrijini kwi-2-8℃ iintsuku ezisi-7 kwaye igcinwe ngaphantsi kwe-15°C iinyanga ezi-6
3.Zonke iisampuli ziphephe imijikelo yokuqandisa nokunyibilikisa.

INKQUBO YOVAVANYO
Nceda ufunde incwadi yemiyalelo yokusebenza kwesixhobo kunye nesihlomelo sephakheji ngaphambi kokuvavanya.
.Esi siphumo sovavanyo sesokubhekisela kwiklinikhi kuphela, akufuneki sisebenze njengesiseko sodwa sokuxilongwa kweklinikhi kunye nonyango, ulawulo lweklinikhi lwezigulane kufuneka luqwalaselwe ngokupheleleyo kunye neempawu zalo, imbali yezonyango, olunye uvavanyo lwelabhoratri, impendulo yonyango, i-epidemiology kunye nolunye ulwazi.
.Le reagent isetyenziswa kuphela kuvavanyo lwe-serum kunye ne-plasma. Isenokungafumani ziphumo zichanekileyo xa isetyenziswa kwezinye iisampuli ezifana namathe nomchamo njl.

IIMPAWU ZOKUSEBENZA

IIREFERENSI
1. I-hepatitis emva kokufakwa kwegazi. Ku: Moore SB, ed. Izifo ezidluliselwa ngegazi ezidluliselwa ngegazi. Alington, VA. Am. Assoc. Iibhanki zegazi, iphe. 53-38.
2.Hansen JH, et al. Ukuphazamiseka kwe-HAMA kwi-Murine Monoclonal Antibody-Based Immunoassays[J].J ye-Clin Immunoassay,1993,16:294-299.
3.Levinson SS. Uhlobo lwee-Antibodies ze-Heterophilic kunye nendima ekuphazamisekeni kwe-Immunoassay[J].J ye-Clin Immunoassay,1992,15:108-114.
4.Alter HJ., Purcell RH, Holland PV, et al. (1978) Iarhente edluliselwayo kwi-hepatitis engeyiyo i-A, engeyiyo i-B. I-Lancet I: 459-463.
5.Choo QL,Weiner AJ, Overby LR, Kuo G, Houghton M. (1990) Intsholongwane yeHepatitis C: eyona nto ibangela intsholongwane yehepatitis engeyiyo i-A, engeyiyo i-B. Br Med Bull 46: 423-441.
6.Engvall E, Perlmann P. (1971) Uvavanyo lwe-enzyme oludityaniswe ne-immunosorbent (ELISA):uvavanyo olusemgangathweni lwe-IgG. Immunochemistry 8:871-874.

IINQOBO EZILINDELEKILEYO

I-HCV-Ab<0.02

Kucetyiswa ukuba ilabhoratri nganye imisele uluhlu lwayo oluqhelekileyo olumele inani lezigulana zayo.

IZIPHUMO ZOVAVANYO NOKUTOLIKWA

• Idatha engentla yimiphumo yovavanyo lwe-HCV-Ab reagent, kwaye kucetyiswa ukuba ilabhoratri nganye imisele uluhlu lwamaxabiso okufumanisa i-HCV-Ab afanelekileyo kuluntu olukulo mmandla. Iziphumo ezingentla zezokujonga kuphela.
• Iziphumo zale ndlela zisebenza kuphela kuluhlu lwereferensi olusekwe kule ndlela, kwaye akukho kuthelekiswa ngokuthe ngqo nezinye iindlela.
• Ezinye izinto zinokubangela iimpazamo kwiziphumo zokufumanisa, kuquka izizathu zobugcisa, iimpazamo zokusebenza kunye nezinye izinto zesampulu.

UKUGCINWA NOKUZINZISA

1. Le khithi ihlala iinyanga ezili-18 ukususela kumhla wokwenziwa kwayo. Gcina ezi khithi zingasetyenziswanga kwi-2-30°C. SUKUZIQANDA. Musa ukuzisebenzisa emva komhla wokuphelelwa.
2. Musa ukuvula isikhwama esivaliweyo ude ulungele ukwenza uvavanyo, kwaye uvavanyo olusetyenziswa kanye lucetyiswa ukuba lusetyenziswe kwindawo efunekayo (ubushushu obuyi-2-35℃, umswakama oyi-40-90%) kwimizuzu engama-60 ngokukhawuleza okukhulu.
3. Isampuli yokuxuba isetyenziswa kwangoko emva kokuba ivuliwe.

IZILUMKISO NEZILUMKISO
.Ikhithi kufuneka ivalwe kwaye ikhuselwe kumswakama.
Zonke iisampulu ezilungileyo maziqinisekiswe ngezinye iindlela.
.Zonke iisampulu maziphathwe njengezingcolisi ezinokubakho.
.MUSA ukusebenzisa i-reagent ephelelwe lixesha.
.MUSA UKUTSHINTSHANA ii-reagents phakathi kweekiti ezinenombolo eyahlukileyo.
.SUKUYISEBENZISA kwakhona amakhadi ovavanyo kunye nazo naziphi na izixhobo ezilahlwayo.
.Ukusetyenziswa gwenxa, isampuli egqithisileyo okanye encinci kunokukhokelela ekuphambukeni kweziphumo.

LUKULINGANISA
.Njengakwezinye iindlela zovavanyo ezisebenzisa ii-antibodies zegundane, kunokwenzeka ukuba kubekho ukuphazamiseka kwee-antibodies ze-human anti-mouse (HAMA) kwisampuli. Iisampuli ezivela kwizigulana ezifumene amalungiselelo e-monoclonal antibodies ukuze zixilongwe okanye zifumane unyango zinokuba ne-HAMA. Ezi sampuli zinokubangela iziphumo ezilungileyo okanye ezimbi.
Isitshixo seempawu ezisetyenzisiweyo:

	Isixhobo Sonyango Sokuxilonga se-In Vitro
	Umenzi
	Gcina kwi-2-30℃
	Umhla WOKUPHELELWA
	Musa Ukuphinda Usebenzise
	ISILUMKISO
	Jonga Imiyalelo Yokusetyenziswa