Kuyesa kwa labotale kumayamba ndi kusonkhanitsa zitsanzo, ndipo kusonkhanitsa zitsanzo ndikosavuta kunyalanyaza.Chofunikira kwambiri pakutolera zitsanzo ndikusankha mtundu wolondola wa zitsanzo, kugwiritsa ntchito zida zoyenera zotsatsira, ndikuyendetsa ndi kukonza bwino.

I. Mtundu wa chitsanzo

Mitundu yodziwika bwino ndi iyi:

Njira yotsatsira sampuli imatha kutanthauza zaukadaulo pakuwunika kwachilengedwe kwa coronavirus yatsopano pamsika wamalonda waulimi (bazaar) ”:

1. Zitsanzo za swab kuchokera ku swabs zapakhosi, manja, zovala ndi zinthu zina za asing'anga: ndikofunikira kugwiritsa ntchito njira yotetezera kachilomboka mu chubu chotengera kachilombo ndikulowetsamo thonje lachitsanzo.Pamwamba pa nthaka) amagwiritsidwa ntchito mobwerezabwereza ndikuchapitsidwa nthawi zopitilira 3.Pa nthawi yomweyo, m'pofunika kuchita Mipikisano mfundo anagawira sampuli pamwamba pa sampuli chinthu.

2. Zitsanzo za swab zazakudya: Zitsanzo za chakudya sizingasonkhanitsidwe mwachindunji.Chakudya chomwe chidzasonkhanitsidwe chizisiyanitsidwa mosamala ndikusungidwa m'thumba lachitsanzo laukhondo musanatenge zitsanzo za swab.M`pofunika kugwiritsa ntchito HIV kuteteza njira mu HIV zitsanzo chubu, pambuyo akuviika mokwanira thonje swab, mobwerezabwereza kupaka ndi muzimutsuka pamwamba pa chakudya chitsanzo kusonkhanitsa kuposa katatu.Pa nthawi yomweyo, m'pofunika kuchita Mipikisano mfundo anagawira sampuli pa chitsanzo pamwamba.

3. Zitsanzo za zimbudzi: Malinga ndi kagawidwe ka ngalande mumsika, sankhani 2-3 zimbudzi sampling malo, kuyang'ana pa mkati chitoliro zosonkhanitsira maukonde, kunsi kwa madzi otaya malangizo, kapena kugwirizana ndi maukonde chitoliro tauni.Kuti mutenge chitsanzo cha swab, ivinizeni m'chimbudzi ndi chitsanzo cha thonje swab kuti ikhale yonyansa ndikutsuka mu chubu lachitsanzo kuposa katatu.Kuti mutenge zitsanzo za zimbudzi, gwiritsani ntchito mabotolo apulasitiki a polyethylene kuti mutenge zitsanzo za madzi a 30-500 mL;Kutolera zimbudzi zokhala ndi voliyumu yopitilira 500 ml, ndowa zapulasitiki za polyethylene kapena zida zapadera zolemeretsa zamadzi zomwe zili pamalopo zingagwiritsidwe ntchito.Panthawi imodzimodziyo, m'pofunika kuchita sampuli zogawidwa zamitundu yambiri za malo osungira zimbudzi.

4. Zitsanzo za nyama: Kwa nyama zamoyo, sampuli za swabs zingagwiritsidwe ntchito kutolera zopota za thupi, zotsekemera za oropharyngeal ndi matako, ndi ndowe kapena zitsanzo za ukazi zikhoza kusonkhanitsidwa ndi kulembedwa moyenerera pa pepala lolembera.Kwa zitsanzo za nyama zomwe zadulidwa zikopa, ndi zina zotero, gwiritsani ntchito swabs za thonje kuti mutenge zitsanzo za thupi lawo ndi thupi lawo.Ndikofunikira kugwiritsa ntchito njira yotetezera kachilomboka mu chubu lachitsanzo la kachilomboka kuti mulowetse munsanja ya thonje, ndikubwerezanso kugwiritsa ntchito pamwamba pazakudya kuti mutenge.Shabu nthawi zopitilira 3.Pa nthawi yomweyo, m'pofunika kuchita Mipikisano mfundo anagawira sampuli pa chitsanzo pamwamba.

5. Zida zina: zotengera zonyamulira ndi kubereketsa nyama monga makola kapena matanki ansomba.Choyamba, yang'anani kapena kumvetsetsa mitundu yeniyeni ya nyama zomwe zasungidwa ndi kuŵetedwa mu chidebe, ndipo sonkhanitsani zitsanzo za swab pakhoma lamkati la chidebecho kapena zitsanzo zamadzimadzi zomwe zili mkati mwake.

6.Sonkhanitsani zitsanzo za aerosol m'madera omwe anthu amasonkhana, malo ochitira malonda am'deralo opanda mpweya wabwino, maofesi, ndi zipinda zopumira.

II.Sampling zida
1. Chubu chotolera magazi
Ngati mukufuna kuyesa nucleic acid ndi zitsanzo za anticoagulant pambuyo pake, ndi bwino kugwiritsa ntchito chotengera cha vacuum chomwe chili ndi EDTA anticoagulant kuti mutenge magazi;Kwa zitsanzo za seramu, tikulimbikitsidwa kugwiritsa ntchito chotengera cha vacuum popanda anticoagulant.

2. Sampling swab

Ndi bwino kugwiritsa ntchito swabs polypropylene m'malo kashiamu alginate swabs kapena swabs ndi mitengo matabwa, chifukwa iwo akhoza kukhala ndi zinthu inactivate mavairasi ena ndi ziletsa PCR kuyezetsa.Nsalu zokhala ndi mitu ya thonje sizingagwiritsidwe ntchito, chifukwa ulusi wa thonje umakhala ndi mphamvu yolimbikitsira mapuloteni ndipo sizimachotsedwa mosavuta mu njira yotsatira yosungira.

Mu "Technical Specifications for 10-in-1 Mixed Collection and Detection of New Coronavirus Nucleic Acids", zanenedwa momveka bwino kuti "sampling swab iyenera kupangidwa ndi poliyesitala, nayiloni ndi zinthu zina zopanda thonje komanso zopanda calcium alginate, ndipo chogwiriracho chiyenera kupangidwa ndi zinthu zopanda matabwa.Malo osweka ndi pafupifupi 3cm kuchokera pamwamba pamutu wa swab, womwe ndi wosavuta kusweka. "

Bungwe la CDC ku United States linanenanso kuti “Gwiritsirani ntchito masinthidwe opangidwa ndi pulasitiki kapena waya.Osagwiritsa ntchito ma swabs a calcium alginate kapena maswabs okhala ndi matabwa,chifukwa amatha kukhala ndi zinthu zomwe zimalepheretsa ma virus ena ndikuletsa kuyesa kwa PCR.

Pakali pano amagwiritsidwa ntchito kwambiri ndi swab yosabala ya nayiloni yosakanizidwa pamodzi ndi ndodo yapulasitiki.Nsapato zoyandama zimakhala ndi mayamwidwe abwinoko amadzi komanso kutulutsa.Malingana ndi malo osiyana siyana, mutu wa swab ulinso ndi makulidwe osiyanasiyana ndi ulusi.Kunenepa.”

3. Zitsanzo zosungirako mayendedwe

Virus inactivation kuteteza njira: mlingo woyenera wa guanidine mchere ndende ndi zosiyanasiyana HIV lysis zigawo zikuluzikulu akhoza mwachindunji lyse kachilombo kumasula nucleic asidi, inactivate RNase ndi DNase, ndi kuonetsetsa umphumphu wa asidi nucleic, amene angagwiritsidwe ntchito qPCR kudziwika wotsatira.Sichingagwiritsidwe ntchito kulima ma virus komanso kudzipatula.

Kuyendera ma virus ndi njira yotetezera: lili ndi mchere, ma amino acid, mavitamini, shuga ndi mapuloteni ofunikira kuti kachilomboka kakhale ndi moyo, zomwe zimatha kusunga ntchito ya kachilomboka.Itha kugwiritsidwa ntchito ngati nucleic acid m'zigawo ndikuzindikira kachilomboka m'tsogolomu, ndipo itha kugwiritsidwanso ntchito kulima ndi kulekanitsa kachilomboka.

4. Chitsanzo chosungira chubu ndi thumba lachitsanzo

Chitsanzo chosungiramo chubu chikuyenera kukhala chubu chapulasitiki chosabala chokhala ndi kapu yopukutira yomwe simatsitsa nucleic acid.Chitsanzo chilichonse chachubu chiyenera kuphimbidwa ndi thumba lachitsanzo kuti chiteteze kutayikira ndi kuipitsidwa kwa chitsanzocho.

III.Kusokoneza zinthu mu chitsanzo

Zinthu zosokoneza pakuyesa kwa PCR

1. amkati kusokoneza

Zinthu zomwe zimapezeka m'thupi zimatha kusokoneza kuzindikira kwa zinthu zina.

Magazi:heme (> 1mg/ml), hemin (> 0.1ng/μl), triglycerides, IgG, etc.;

Mkodzo:Urea (20mM);

Choponda:mbewu za polysaccharides, cholate;

Thupi:mapuloteni, collagen;

Mkaka: mapuloteni, calcium ion;

Mankhwala osokoneza bongo: maantibayotiki, mankhwala oletsa tizilombo toyambitsa matenda, mahomoni, ndi zina zotero.

zina.

2. kusokoneza kunja

Zimachokera ku zinthu zomwe zingasokoneze kudziwika kwa zinthu zina mu vitro.Zitsanzo zowonjezera ndi zinthu zomwe zitha kulumikizidwa pakusonkhanitsa ndi kukonza zitsanzo:

Zida zoyesera: gel osakaniza seramu, chidebe chosonkhanitsira zitsanzo ndi choyimitsa mphira, catheter, catheter flushing fluid, gulovu ufa, etc.

Anticoagulant: heparin (> 0.1 U / mL), EDTA (> 0.5 mM), etc.;

organic solvents: isopropanol (> 0.1 IU/ml), betaine, dimethyl sulfoxide, formamide, glycerin, PEG, etc.;

Mankhwala opha tizilombo: mowa (ethanol> 1%), aldehydes, phenol (phenol> 0.2%), peroxides, amphamvu zidulo (HCL), alkalis amphamvu (NaOH), etc.;

Chotsukira:SDS (> 0.005%);

Nthaka: humus

Dyestuff: Indigo

zina.

Pansi pazitsanzo zenizeni zakunja, monga kusonkhanitsa zitsanzo zachilengedwe, zitsanzo zimatha kukhala ndi zinthu zosiyanasiyana zosokoneza, ndipo mapangidwe a zinthu zosokoneza m'malo aliwonse ndi osiyana, zomwe zimafuna kuyesa kusokoneza kwa PCR kuti kuchitidwe malinga ndi momwe zinthu zilili.

IV.Zitsanzo zosungirako ndi zoyendera

Zitsanzo zachipatala zomwe zasonkhanitsidwa kumene ziyenera kutumizidwa ku labotale mkati mwa maola 2 mpaka 4 mutatoledwa pa 2°C mpaka 8°C.Sungani pa 4 ℃ mkati mwa 24h.Zitsanzo zomwe zimagwiritsidwa ntchito podzipatula ma virus komanso kuzindikira kwa nucleic acid ziyenera kuyesedwa posachedwa.Zitsanzo zomwe zingathe kuyesedwa mkati mwa maola 24 zikhoza kusungidwa pa 4 ° C;zitsanzo zomwe sizingawoneke mkati mwa maola 24 ziyenera kusungidwa pa -70 ° C kapena pansi (ngati sichoncho- Sungani pa 70 ° C mufiriji pa -20 ° C).

Chidule:Kutolera kokhazikika, kunyamula ndi kusungirako zitsanzo ndizofunikira pakuyesedwa kotsatira.Zoyipa zabodza zamayeso ambiri a labotale zimayamba chifukwa chosonkhanitsira molakwika, kunyamula ndi kusungirako zitsanzo zomwe zimapangitsa kuwonongeka kwa ma virus nucleic acid.Komabe, ma laboratories ambiri amanyalanyazabe kufunikira kwa zitsanzo.Ndikukhulupirira kuti nkhaniyi ikhoza kudzutsa chidwi cha aliyense pakupanga zitsanzo ndikupanga zitsanzo zasayansi komanso zomveka!

Zogwirizana nazo:

SARS-CoV-2 Variant Nucleic Acid Detection Kit (I) (Multiplex PCR Fluorescent Probe Method) -Kuzindikira Ma Mutants ochokera ku Brazil, South Africa, ndi UK