Nnwale ụlọ nyocha na-amalite site na nchịkọta nlele, na nchịkọta sample bụ nke kachasị mfe ileghara anya.Ihe kachasị mkpa maka nchịkọta nlele bụ ịhọrọ ụdị nlele ziri ezi, jiri ngwaọrụ nlele kwesịrị ekwesị, na-ebu njem na nhazi ezi uche dị na ya.

I.Sample ụdị

Ụdị ihe atụ a na-ahụkarị bụ ndị a:

Usoro nlele nwere ike na-ezo aka na nkọwapụta teknụzụ maka nlekota gburugburu ebe obibi nke coronavirus ọhụrụ n'ahịa ahia ugbo (bazaar)":

1. Swab samples site na akpịrị akpịrị, aka, uwe na ihe ndị ọzọ nke ndị na-eme ya: a chọrọ iji ihe nchekwa nchekwa nje na tube nlele nje wee banye n'ụzọ zuru ezu na swabs owu.A na-etinye elu ala) ugboro ugboro ma kpochaa ihe karịrị ugboro 3.N'otu oge ahụ, ọ dị mkpa ka a na-eme ọtụtụ ihe na-ekesa ihe nlele n'elu ihe atụpụtara.

2. Nri nri elu swab samples: nri nri enweghị ike ịnakọta ozugbo.Ekwesịrị iji nlezianya kewaa nri a ga-anakọta ma debe ya n'ime akpa nlele dị ọcha tupu ịnakọta swab sample.A na-achọrọ iji ihe nchekwa nchekwa nje dị na tube nlele nje virus, mgbe ọ gbasasịrị nke ọma n'ụdị swab owu, ka a na-ete ya ugboro ugboro ma sachaa elu nke ihe oriri a ga-anakọta ihe karịrị ugboro atọ.N'otu oge ahụ, ọ dị mkpa ka a na-eme ọtụtụ ihe na-ekesa ihe nlele n'elu ihe atụ.

3. Ihe nlele nke mmiri mmiri: Dị ka nkesa nke usoro mmiri ozuzo na ahịa, họrọ 2-3 ebe a na-enyocha mmiri mmiri, na-elekwasị anya na nchịkọta netwọk nke ime ụlọ, n'okpuru mmiri nke mmiri na-asọba, ma ọ bụ njikọ na netwọk ọkpọkọ obodo.Iji nakọta ihe nlele swab, gbanye ya n'ime mmiri mmiri na-eji owu swab sample iji mee ka ọ gbasaa nsị ahụ ma kpochaa ya na tube nlele ihe karịrị ugboro atọ.Iji nakọta ihe nlele nsị, jiri karama plastik polyethylene na-anakọta ihe nlele mmiri nsị 30-500 ml;maka nchịkọta nsị nke nwere oke karịa 500 ml, ị nwere ike iji bọket plastik polyethylene ma ọ bụ ihe nlele mmiri dị na saịtị ahụ nwere ike iji akụrụngwa nkwalite pụrụ iche.N'otu oge ahụ, ọ dị mkpa ka ị na-eduzi nleba anya nkesa ọtụtụ ebe maka ebe nlele nsị.

4. Anụmanụ anụmanụ: Maka anụmanụ dị ndụ, a pụrụ iji swabs swabs na-anakọta swabs elu ahụ, oropharyngeal swabs na anal swabs, na nsị ma ọ bụ ihe nzuzo nwere ike ịnakọta ma dekọọ ya na akwụkwọ ndekọ.Maka ihe nlele anụmanụ nke megoro akpụkpọ anụ, wdg, jiri swabs owu na-anakọta elu ahụ ha na oghere swab nke oghere ahụ.A na-achọrọ iji ihe nchekwa nchekwa nje dị na tube nlele nje iji banye n'ime akwa swab nke sampling, wee megharịa ngwa n'elu ihe nri a ga-anakọta.Shabu karịa ugboro 3.N'otu oge ahụ, ọ dị mkpa ka a na-eme ọtụtụ ihe na-ekesa ihe nlele n'elu ihe atụ.

5. Ngwa ndị ọzọ: igbe maka ibufe na ịzụ anụmanụ dị ka ngịga ma ọ bụ tank azụ.Nke mbụ, lelee ma ọ bụ ghọta ụdị anụmanụ ndị echekwara na bred n'ime akpa ahụ, ma na-anakọta swab n'ime mgbidi dị n'ime akpa ahụ ma ọ bụ ihe nlele mmiri dị n'ime ya.

6.Collect aerosol samples na mpaghara ebe ndị mmadụ na-ezukọ, ikuku ikuku na-adịghị mma na mpaghara azụmahịa, ụlọ ọrụ, na ụlọ ezumike.

II.Nlele ngwaọrụ
1. Ọbara nchịkọta tube
Ọ bụrụ na ị ga-eme nyocha nucleic acid na ihe nleba anya na-egbu egbu ma emechaa, a na-atụ aro ka ị jiri arịa ọbara na-ekpo ọkụ nwere EDTA anticoagulant iji nakọta ọbara;maka ihe nlele ọbara, a na-atụ aro ka ị jiri arịa ọbara na-ekpo ọkụ na-enweghị ọgwụ mgbochi.

2. Nlereanya swab

A na-atụ aro ka iji polypropylene swabs kama calcium alginate swabs ma ọ bụ swabs nwere ogwe osisi, n'ihi na ha nwere ike ịnwe ihe ndị na-adịghị arụ ọrụ ụfọdụ nje ma gbochie ule PCR.Enweghị ike iji swabs nwere isi owu, n'ihi na eriri owu nwere nnukwu mgbasa ozi nke protein na ọ naghị adị mfe ịbanye na ngwọta nchekwa na-esote.

Na "Nkọwa nka nka maka mkpokọta 10-in-1 Mixed and Detection of New Coronavirus Nucleic Acids", a na-akọwa n'ụzọ doro anya na "a ga-eme swab sampling nke polyester, naịlọn na ihe ndị ọzọ na-abụghị owu na nke na-abụghị calcium alginate, na aka kwesịrị ka ọ bụrụ ihe na-abụghị osisi.Ebe mgbawa dị ihe dịka 3cm site n'elu isi swab, nke dị mfe ịgbaji. "

CDC dị na United States kwukwara “Jiri naanị swabs sịntetik nke nwere rọba ma ọ bụ osisi waya.Ejila calcium alginate swabs ma ọ bụ swabs nwere ogwe osisi,dịka ha nwere ike ịnwe ihe na-eme ka ụfọdụ nje rụọ ọrụ ma gbochie ule PCR.

A na-ejikarị eme ihe ugbu a bụ swab nylon na-adịghị mma nke a na-ekpofu ya yana mkpanaka rọba.Igwe swab na-akpụ akpụ na-enwe nnabata mmiri ka mma yana ọnụego ntọhapụ.Dịka saịtị nlele dị iche iche si dị, isi swab nwekwara nha na eriri dị iche iche.The ọkpụrụkpụ.”

3. Ihe ngwọta nchekwa njem ihe atụ

Ihe ngwọta nchekwa inactivation nke nje: ọnụọgụ nnu guanidine kwesịrị ekwesị na ihe dị iche iche nke nje virus nwere ike lyse nje ahụ ka ọ hapụ acid nucleic, na-adịghị arụ ọrụ RNase na DNase, ma hụ na iguzosi ike n'ezi ihe nke nucleic acid, nke enwere ike iji mee nchọpụta qPCR na-esote.Enweghị ike iji ya mee ihe maka ịkọ nje na ikewapụ ya.

Njefe njem na nchekwa ihe ngwọta: nwere salts, amino acids, vitamin, glucose na protein dị mkpa maka nlanarị nke nje, nke nwere ike ịnọgide na-arụ ọrụ nke nje.Enwere ike iji ya mee ihe maka mmịpụta nucleic acid na nchọpụta nje n'ọdịnihu, a pụkwara iji ya mee ihe maka ịkụ ihe na nkewa nke nje ahụ.

4. tube nchekwa ihe nlele na akpa nlele

Igwe nchekwa ihe nlele kwesịrị ịbụ tube rọba na-adịghị ọcha nke nwere okpu ịghasa nke na-adịghị etinye nucleic acid.Ekwesịrị iji akpa nlele kpuchie tube ọ bụla iji gbochie ntapu na mmetọ nke ihe nlele ahụ.

III.Ihe ntinye aka na ihe nlele

Ihe ntinye aka na nnwale PCR

1. endogenous nnyonye anya

Ihe ndị dị na nlele ahụ nwere ike igbochi ịchọpụta ihe ndị ọzọ.

Ọbara:heme (> 1 mg / ml), hemin (> 0.1ng / μl), triglycerides, IgG, wdg;

Urine:Urea (20mM);

stool:osisi polysaccharides, cholate;

Anụ ahụ:protein, collagen;

Mmiri ara: protease, calcium ion;

Ọgwụ ọjọọ: ọgwụ nje, ọgwụ mgbochi nje, homonụ, wdg.

ọzọ.

2. exogenous nnyonye anya

Ọ na-abịa site na ihe ndị nwere ike igbochi nchọpụta nke ihe ndị ọzọ na vitro.Ihe mgbakwunye ihe atụ na ihe enwere ike ịkpọtụrụ n'oge nchịkọta na nhazi nlele:

Nnwale akụrụngwa: serum separation gel, sample collection container and roba stopper, catheter, catheter flushing fluid, glove powder, etc.

Ọgwụ mgbochi: heparin (> 0.1 U / ml), EDTA (> 0.5 mM), wdg;

Ihe mgbaze organic: isopropanol (> 0.1 IU / ml), betaine, dimethyl sulfoxide, foramide, glycerin, PEG, wdg;

Ọgwụ nje: alcohols (ethanol> 1%), aldehydes, phenols (phenol> 0.2%), peroxides, acid siri ike (HCL), alkalis siri ike (NaOH), wdg;

Ngwa ncha:SDS (> 0.005%);

Ala: humus

Ihe eji eme ihe: Indigo

ọzọ.

N'okpuru ọnọdụ nlele n'èzí n'ezie, dị ka ịnakọta ihe atụ gburugburu ebe obibi, ihe nlele ahụ nwere ike ịnwe ihe dị iche iche na-etinye aka na ya, na ihe mejupụtara ihe ndị na-etinye aka na ebe ọ bụla dị iche iche, nke chọrọ nyocha nnyonye anya PCR dị ka ọnọdụ mpaghara si dị.

IV.Ihe nlele nchekwa na ọnọdụ njem

Ekwesịrị iziga ụdị ọgwụgwọ ekpokọtara ọhụrụ n'ụlọ nyocha n'ime awa 2 ruo 4 mgbe anakọtara na 2 Celsius ruo 8 Celsius.Chekwaa na 4 ℃ n'ime awa 24.A ga-anwale ụdịdị eji ekewapụ nje virus na nchọpụta nucleic acid ozugbo enwere ike.Enwere ike idobe ihe nlele nke enwere ike ịnwale n'ime awa 24 na 4 Celsius;A ga-echekwa ụdịdị a na-agaghị achọpụta n'ime awa 24 na -70C ma ọ bụ n'okpuru (ọ bụrụ na ọ bụghị - Chekwaa na 70°C n'ime friji na -20°C).

Nchịkọta:Nchịkọta ahaziri ahazi, njem na nchekwa nke ihe nlele dị mkpa maka ule na-esote.Ihe na-adịghị mma nke ọtụtụ ule ụlọ nyocha na-akpata site na nchịkọta na-ezighị ezi, njem na nchekwa nke ihe nlele nke na-eduga na mmebi nke nje nucleic acid.Agbanyeghị, ọtụtụ ụlọ nyocha ka na-eleghara mkpa nlele anya.Enwere m olileanya na akụkọ a nwere ike ịkpọlite ​​uche onye ọ bụla na nlele ma mee ka nyocha sayensị na ezi uche dị ukwuu!

Ngwaahịa emetụtara:

Ngwa SARS-CoV-2 dị iche iche Nchọpụta Nucleic Acid (I) (Usoro nyocha nke ọtụtụ PCR Fluorescent) - Maka nchọpụta nke ụmụ anụmanụ sitere na Brazil, South Africa na UK