Kuita kweina nucleic acid amplification assays kuona SARS-CoV-2 muEthiopia

Ndatenda nekushanyira Nature.com. Uri kushandisa shanduro yebrowser ine tsigiro shoma yeCSS. Kuti uwane ruzivo rwakanyanya, tinokurudzira kuti ushandise browser yakagadziridzwa (kana kudzima Compatibility Mode muInternet Explorer). Pamusoro pezvo, kuti tive nechokwadi chekutsigira kunoenderera, tinoratidza saiti isina masitaera uye JavaScript.
Inoratidza carousel yemasiraidhi matatu panguva imwe chete. Shandisa mabhatani ePashure neAnotevera kufamba nemasiraidhi matatu panguva, kana kushandisa mabhatani anotsveyamisa ari kumagumo kufamba nemasiraidhi matatu panguva.
Kubva muna 2019 chirwere chekoronavirus (COVID-19) chatanga, akawanda ekutengesa nucleic acid amplification bvunzo (NAATs) akagadzirwa kutenderera pasirese uye ave assays akajairwa. Kunyange zvazvo bvunzo dzinoverengeka dzakakurumidza kugadzirwa uye dzakashandiswa kumarabhoritari diagnostic bvunzo, kuita kwebvunzo idzi hakuna kuongororwa munzvimbo dzakasiyana. Naizvozvo, chidzidzo ichi chine chinangwa chekuongorora mashandiro eAbbott SARS-CoV-2, Daan Gene, BGI, uye Sansure Biotech assays vachishandisa Composite Reference Standard (CRS). Chidzidzo chacho chakaitwa paEthiopian Public Health Institute (EPHI) kubva 1 kusvika 30 December 2020. 164 nasopharyngeal samples dzakatorwa uchishandisa QIAamp RNA mini kit uye Abbott DNA samples preparation system. Pazviyero zana nemakumi matanhatu neina, 59.1% yaive yakanaka uye 40.9% yaive isina kunaka kune CRS. Sansure Biotech positivity yaive yakaderera zvakanyanya kana ichienzaniswa neCRS (p <0.05). Sansure Biotech positivity yaive yakaderera zvakanyanya kana ichienzaniswa neCRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p <0,05). Mhedzisiro yakanaka yeSansure Biotech yaive yakaderera zvakanyanya zvichienzaniswa neCRS (p <0.05).与CRS 相比，Sansure Biotech 的阳性率显着较低（p <0.05）.与CRS 相比，Sansure Biotech 的阳性率显着较低（p <0.05）. У Sansure Biotech было значительно меньше положительных результатов по сравнению с CRS (p <0,05). Sansure Biotech yaive nemhedzisiro shoma yakanaka kana ichienzaniswa neCRS (p <0.05).Chibvumirano chese chekuongorora ina chaive 96.3-100% zvichienzaniswa neCRS. Pamusoro peiyo yakaderera positivity mwero weSansure Biotech assay, kuita kwemaassays mana kwakada kufananidzwa. Saizvozvo, iyo Sansure Biotech [Research Chete (RUO)] bvunzo inoda kumwe kusimbiswa kwekushandiswa kwayo muEthiopia. Chekupedzisira, tsvakiridzo yekuwedzera inofanirwa kutariswa kuti iongorore ma assays ane akakodzera zvirevo zvemugadziri.
Kuongororwa murabhoritari chikamu cheWorld Health Organisation (WHO) Strategic Plan yeCoronavirus Chirwere 2019 (COVID-19) Preparedness and Response (SPRP). WHO inopa zano kuti nyika dzinoda kuvaka marabhoritari hunyanzvi hwekuvandudza kugadzirira, kutonga kwakaringana, kusvinura uye kupindura nekukurumidza kumatambudziko ehutano hweveruzhinji. Izvi zvinoratidza kuti basa rerabhoritari rakakosha kuratidza chirwere uye epidemiology yezvinotapukira zvinotapukira uye kudzora kupararira kwavo.
Kuongororwa kweCCIDID-19 kunoda epidemiological uye ruzivo rwekurapa, zviratidzo zvemunhu / zviratidzo, uye radiographic nerabhoritari data. Kubva pakabuda COVID-19 muWuhan, China, akawanda ekutengesa nucleic acid amplification bvunzo (NAATs) akagadzirwa pasirese. Real-time reverse transcription polymerase chain reaction (rRT-PCR) yakashandiswa senzira uye yakajairwa nzira yekuongororwa murabhoritari yeyakakomba acute respiratory syndrome 2 (SARS-CoV-2)3 utachiona. Kuonekwa kwemamorekuru eSARS-CoV-2 inowanzoenderana neN (nucleocapsid protein gene), E (envelope protein gene), uye RdRp (RNA-inotsamira RNA polymerase gene) muORF1a/b (yakavhurika yekuverenga furemu 1a/b) . gene) dunhu rinozivikanwa kubva kuhutachiona genome. Iwo anoonekwa seakanyanya kuchengetedzwa matunhu anowanikwa muhutachiona genomes yekuzivikanwa kwehutachiona4. Pakati peaya majini, iyo RdRp neE majini ane yakakwirira analytical yekuona senitivity, nepo N gene ine yakaderera analytical senitivity5.
Kuita kwePCR assays kunogona kusiyana zvichienderana nezvinhu zvakasiyana-siyana se: mareji ekubvisa, amplification / detection reagents, nzira yekubvisa, kunaka kwePCR muchina uye zvimwe zviridzwa. Kubva muna Kubvumbi 2020, anopfuura makumi mana nemasere ekuongorora akasiyana kubva kunyika pfumbamwe akagamuchira Emergency Use Authorization (EUA) yeCCIDID-196 diagnostics. MuEthiopia, anopfuura gumi nemana chaiwo-nguva PCR mapuratifomu anoshandiswa kuona PCR yeSARS-CoV-2 pamakumi maviri nenhanhatu hutano masangano, kusanganisira ABI 7500, Abbott m2000, Roche 48000 uye Quant-studio7. Uye zvakare, akasiyana PCR bvunzo kits anowanikwa, akadai seDaan Gene bvunzo, Abbott SARS-CoV-2 bvunzo, Sansure Biotech bvunzo, uye SARS-CoV-2 BGI bvunzo. Kunyangwe rRT-PCR ine hanya zvakanyanya, vamwe varwere vane COVID-19 vanoshuma mhedzisiro yakaipa nekuda kwekusakwana makopi eviral ribonucleic acid (RNA) mumasample nekuda kwekuunganidza zvisirizvo, kutakura, kuchengetedza uye kubata, uye kuyedza marabhoritari. mamiriro nezviito zvevashandi8. Pamusoro pezvo, sampuli kana kudzora kubata zvisina kunaka, kutenderera chikumbaridzo (Ct) kuseta, uye kuyambuka-reactivity nemamwe pathogenic nucleic acids kana kusashanda/kusara SARS-CoV-2 RNA kunogona kutungamirira kune nhema dzakanaka mhedzisiro murRT-PCR9 assays. Saka, zviri pachena kuti PCR bvunzo dzinogona zvechokwadi kuona vatakuri vezvimedu zvejeni, sezvo vasingatombokwanise kusiyanisa pakati peanoshanda chaiwo majeni ehutachiona, saka bvunzo dzinogona kungoziva vatakuri uye kwete varwere10. Naizvozvo, zvakakosha kuti tiongorore mashandiro ekuongorora tichishandisa nzira dzakajairwa mugadziriro yedu. Kunyangwe mareagents mazhinji eNAAT achiwanikwa kuEthiopian Public Health Institute (EPHI) uye nemunyika yese, hapana ongororo yekuenzanisa yekushanda kwavo isati yataurwa. Naizvozvo, chidzidzo ichi chine chinangwa chekuongorora kuenzaniswa kwematekisheni anowanikwa mune zvekutengesa kuti aonekwe kweSARS-CoV-2 ne rRT-PCR uchishandisa sampuli dzekiriniki.
Vatori vechikamu 164 vane vanofungidzirwa kuti COVID-19 vakaiswa muchidzidzo ichi. Ruzhinji rwemasamples aibva kunzvimbo dzekurapa (118/164 = 72%), nepo vasara makumi mana nematanhatu (28%) vatori vechikamu vaibva kunzvimbo dzisiri dzekurapa. Pakati pevatori vechikamu vasina kurapwa pamuzinda, gumi nemashanu (9.1%) vaive nemhosva dzekufungidzirwa uye 31 (18.9%) vaive nevanobatika nenyaya dzakasimbiswa. Makumi mapfumbamwe nevatatu (56.7%) vatori vechikamu vaive varume, uye zvinoreva (± SD) zera revatori vechikamu raive 31.10 (± 11.82) makore.
Muchidzidzo ichi, mwero wakanaka uye wakashata webvunzo ina dzeCOVID-19 dzakatemwa. Saka, mwero wakanaka weAbbott SARS-CoV-2 assay, Daan Gene 2019-nCoV assay, SARS-CoV-2 BGI assay, uye Sansure Biotech 2019-nCoV assay yaive 59.1%, 58.5%, 57.9% uye 55.5% zvakateerana. . Izvo zvakanaka uye zvisina kunaka composite reference standard (CRS) zvibodzwa zvaive 97 (59.1%) uye 67 (40.9%), zvichiteerana (Tafura 1). Muchidzidzo ichi, tsananguro yeCRS yaive yakavakirwa pamutemo we "chero wakanaka", wekuti kubva pamiyedzo ina, maviri kana anopfuura ebvunzo mhinduro akapa mhedzisiro yakafanana akaonekwa seakanaka kana asina kunaka.
Muchidzidzo ichi, takawana chibvumirano chakashata chezana (NPA) che100% (95% CI 94.6-100) chekuongorora kwese kuchienzaniswa neCRS. Iyo Sansure Biotechnology ongororo yakaratidza shoma PPA ye93.8% (95% CI 87.2-97.1) uye iyo Daan Gene 2019-nCoV ongororo yaive nechibvumirano chakazara che99.4% (95% CI 96.6-99.9). Kusiyana neizvi, chibvumirano chakazara pakati peSARS-CoV-2 BGI assay uye Sansure Biotech 2019-nCoV assay yaive 98.8% uye 96.3%, zvichiteerana (Tafura 2).
Cohen's kappa coefficient yechibvumirano pakati peCRS neAbbott SARS-CoV-2 assay mhedzisiro yainyatsoenderana (K = 1.00). Saizvozvo, Cohen's kappa tsika dzakaonekwa naDaan Gene 2019-nCoV, SARS-CoV-2 BGI, uye Sansure Biotech 2019-nCoV zvakare inoenderana zvizere neCRS (K ≥ 0.925). Mukuongorora uku kwekuenzanisa, iyo chi-square bvunzo (McNemar bvunzo) yakaratidza kuti Sansure Biotech 2019-nCoV assay mhinduro dzaive dzakasiyana zvakanyanya neCRS mhedzisiro (p = 0.031) (Tafura 2).
Sezvinoratidzwa muFig.1 iyo muzana yeCt yakaderera kukosha (<20 Ct) yeAbbott SARS-CoV-2 assay (yakasanganiswa RdRp neN gene) yaive 87.6% uye ORF1a/b gene Ct kukosha kweSansure Biotech 2019-nCoV assay yakaratidza kuti chikamu cheyakaderera. Ct kukosha (<20 Ct) yaiva 50.3% uye yakakwirira Ct kukosha (36-40 Ct) yaiva 3.2%. 1 iyo muzana yeCt yakaderera kukosha (<20 Ct) yeAbbott SARS-CoV-2 assay (yakasanganiswa RdRp neN gene) yaive 87.6% uye ORF1a/b gene Ct kukosha kweSansure Biotech 2019-nCoV assay yakaratidza kuti chikamu cheyakaderera. Ct kukosha (<20 Ct) yaiva 50.3% uye yakakwirira Ct kukosha (36-40 Ct) yaiva 3.2%.Sezvinoratidzwa muFig.1, процент наименьшего значения Ct (< 20 Ct) анализа Abbott SARS-CoV-2 (комбинированный ген RdRp и N) составил 87,6%, а значзаение ORF Santech 2019-nCoV показало что процент низкого значения Ct (< 20 Ct) составлял 50,3%, а высокое значение Ct (36–40 Ct) составляло 3,2%. 1, iyo muzana yeyakaderera Ct kukosha (<20 Ct) kuongororwa kweAbbott SARS-CoV-2 (yakasanganiswa gene RdRp uye N) yaive 87.6%, uye kukosha kweCt yeORF1a/b gene kuongororwa kweSansure Biotech 2019-nCoV yakaratidza. kuti muzana weiyo yakaderera Ct kukosha (<20 Ct) yakaverengera 50.3%, uye yakakwirira kukosha Ct (36–40 Ct) yakaverengerwa 3.2%.如图1 所示，Abbott SARS-CoV-2 检测（结合RdRp 和N 基因）的最低Ct 值百分比（< 20 Ct）为19-Biotechnik 87.6%2, San 197.6 %检测的ORF1a/b 基因Ct 值显示低Ct 值(< 20 Ct) 的百分比為50.3%，高Ct 值(36–40 Ct) 的百分比为3. Sezvinoratidzwa mumufananidzo 1, iyo yakaderera Ct kukosha muzana (<20 Ct) yeAbbott SARS-CoV-2 bvunzo (musanganiswa weRdRp neN gene) i87.6%, iyo ORF1a/b gene Ct kukosha kweSansure Biotech 2019-nCoV bvunzo. inoratidza yakaderera Ct 值(<20 Ct) 的 percentage is 50.3%,高Ct 值(36–40 Ct) 的 percentage is 3.2%. Как показано на рисунке 1, анализ Abbott SARS-CoV-2 (сочетающий гены RdRp и N) имел самое низкое процентное значение значение Ct (< 20 Ct% , 7а змени) , 8 гена ORF1a/b в исследовании Sansure Biotech 2019- Анализ nCoV показал низкий Ct. Sezvinoratidzwa mumufananidzo 1, iyo Abbott SARS-CoV-2 assay (kusanganisa iyo RdRp neN gene) yaive neiyo yakaderera muzana muzana Ct kukosha (<20 Ct) pa87.6%, nepo Ct kukosha kweORF1a/b gene muSansure. Biotech 2019 kudzidza - Ongororo yenCoV yakaratidza yakaderera Ct. Процент значений (< 20 Ct) составил 50,3%, а процент высоких значений Ct (36–40 Ct) составил 3,2%. Iwo muzana yezvakakosha (<20 Ct) yaive 50.3%, uye iyo muzana yeyakakwira Ct kukosha (36-40 Ct) yaive 3.2%.Iyo Abbott SARS-CoV-2 B bvunzo yakanyorwa Ct kukosha pamusoro pemakumi matatu. Kune rumwe rutivi, paBGI SARS-CoV-2 assay ORF1a / b gene yaiva nepamusoro Ct kukosha (> 36 Ct) muzana yaiva 4% (Fig. 1). Kune rumwe rutivi, paBGI SARS-CoV-2 assay ORF1a / b gene yaiva nepamusoro Ct kukosha (> 36 Ct) muzana yaiva 4% (Fig. 1). С другой стороны, в анализе BGI SARS-CoV-2 ген ORF1a/b имел высокое значение Ct (> 36 Ct), процент которого составлял 4% (рис. 1). Kune rumwe rutivi, mukuongororwa kweBGI SARS-CoV-2 gene ORF1a / b yakanga ine Ct yakakwira kukosha (> 36 Ct), iyo muzana yaive 4% (Fig. 1).另一方面，在BGI SARS-CoV-2 检测中，ORF1a/b 基因具有高Ct 值（> 36 Ct）的百分比为4%（图1）. Nekune rimwe divi, mukuona kweBGI SARS-CoV-2, iyo percentage yeORF1a/b gene ine high Ct value (>36 Ct) i4% (Figure 1). С другой стороны, в анализе BGI SARS-CoV-2 процент генов ORF1a/b с высокими значениями Ct (>36 Ct) составил 4% (рис. 1). Kune rimwe divi, muBGI SARS-CoV-2 ongororo, iyo muzana yeORF1a/b genes ine yakakwira Ct tsika (> 36 Ct) yaive 4% (Fig. 1).
Muchidzidzo ichi, takatora 164 nasopharyngeal samples. Kune ese marudzi emaassays, kuparadzaniswa kweRNA uye kukwidziridzwa kwakaitwa pachishandiswa nzira nemakiti anokurudzirwa nevagadziri vacho.
Chidzidzo ichi chakaratidza kuti bvunzo yaAbbott yeSARS-CoV-2 ine maitiro akafanana ekuona seCRS, iine 100% yakanaka, yakaipa, uye yakazara concordance. Cohen's kappa chibvumirano ndeye 1.00, zvichiratidza kubvumirana kuzere neCRS. Ongororo yakafanana yakaitwa neYunivhesiti yeWashington kuUS yakawana kuti kunzwisiswa kwese uye kujeka kwebvunzo yeAbbott yeSARS-CoV-2 yaive 93% ne100%, zvichiteerana, zvichienzaniswa nerabhoritari-yakatemerwa assay (LDA) yeCDC. . 11. Iyo Abbott SARS-CoV-2 yekuona sisitimu yakavakirwa pakuona panguva imwe chete yakasanganiswa yeN uye RdRp genes, sezvo majini ese ari maviri anonyanya kunzwisiswa, achideredza nhema dzisina kunaka12. Ongororo muVienna, Austria yakaratidzawo kuti mavhoriyamu akakura ekubvisa sampuli uye kuona mavhoriyamu eeluent akadzikisa dilution mhedzisiro uye kuwedzera kwekuona zvakanaka13. Nekudaro, mutambo waAbbott wakakwana weiyo SARS-CoV-2 assay inogona kubatanidzwa nepuratifomu yekuona sisitimu iyo panguva imwe chete inoona combinatorial majini, inobvisa huwandu hukuru hwemasamples (0.5 ml), uye inoshandisa yakawanda yakawanda (40 µl).
Zvigumisiro zvedu zvakaratidzawo kuti kuonekwa kwekuita kweDaan genetic test kwaive kwakangofanana neCRS. Izvi zvinoenderana nechidzidzo14 chakaitwa paAnhui University muHuainan, China, uye zvakataurwa nemugadziri we100% chibvumirano chakanaka. Kunyangwe mishumo yemhedzisiro inowirirana, imwe sampu yanga isiriyo mushure mekudzokorora zvakafanana eeate, asi yaive yakanaka muAbbott SARS-CoV-2 uye Sansure Biotech nCoV-2019 assays. Izvi zvinoratidza kuti panogona kunge paine mutsauko mumhedzisiro mumhando dzakasiyana dzemayedzo. Zvakangodaro, muchidzidzo chakaitwa muChina15, mhedzisiro yeDaan Gene assay yakanga yakasiyana zvakanyanya (p <0.05) kana ichienzaniswa neyavo-yakatsanangurwa referensi referensi. Zvakangodaro, muchidzidzo chakaitwa muChina15, mhedzisiro yeDaan Gene assay yakanga yakasiyana zvakanyanya (p <0.05) kana ichienzaniswa neyavo-yakatsanangurwa referensi referensi. Тем не менее, в исследовании, проведенном в Китае15, результат анализа Daan Gene значительно отличался (p <0,05) от их лаборанались отличался. Nekudaro, muchidzidzo muChina15, mhedzisiro yekuongorora yaDaan Gene yaive yakasiyana zvakanyanya (p <0.05) kubva kuongororo yavo yerabhoritari.然而，在中国进行的研究中15，大安基因检测的结果与其实验室定义的参考检测相比有显着。）然而，在中国进行的研究中15，大安基因检测的结果与其实验室定义的参考检测相比有显着0. Однако в исследовании, проведенном в Китае15, результаты генетического теста Daan значительно отличались (p <0,05) по сравнетического теста Daan тестом. Nekudaro, mune chidzidzo muChina15, mhedzisiro yeDaan's genetic test yaive yakasiyana zvakanyanya (p <0.05) kana ichienzaniswa neyayo referensi laboratory test.Kusiyana uku kunogona kunge kuri nekuda kwekunzwa kwereferensi bvunzo yekuona SARS-CoV-2, uye zvimwe zvidzidzo zvinogona kuve zvakakosha kuti uone chikonzero.
Pamusoro pezvo, chidzidzo chedu chakaongorora kuenzanisa kwekuita kweSARS-CoV-2 BGI assay neCRS, ichiratidza chibvumirano chakanaka chezana (PPA = 97.9%), chibvumirano chisina kunaka chezana (NPA = 100%), uye chibvumirano chesero chese OPA). ) = 98.8%). Cohen's Kappa tsika dzakaratidza chibvumirano chakanaka (K = 0.975). Zvidzidzo muNetherlands16 neChina15 zvakaratidza mhedzisiro inowirirana. Iyo SARS-CoV-2 BGI bvunzo ijini rimwe chete (ORF1a/b) yekuona bvunzo uchishandisa gumi µl amplification/detection eluate. Pasinei nekubvumirana kwakanaka kwenhamba nemigumisiro yedu yereferensi, ongororo yakapotsa masampuli maviri akanaka (1.22%) yemuenzaniso wakazara. Izvi zvinogona kuve nemhedzisiro yakakura yekiriniki yekutapurirana masimba kune ese murwere uye munharaunda.
Imwe ongororo yekuenzanisa inosanganisirwa muchidzidzo ichi yaive Sansure Biotech nCoV-2019 rRT-PCR (RUO) assay; chikamu chose chemutambo chaive 96.3%. Simba rekubvumirana rakagadziriswawo neCohen's Kappa value, iyo yaiva 0.925, ichiratidza kubvumirana kwakakwana neCRS. Zvakare, mhedzisiro yedu yakafanana nezvidzidzo zvakaitwa kuCentral South University muChangsha, China, uye kuClinical Laboratory Dhipatimendi reLiuzhou People's Hospital, Liuzhou City, China17. Kunyangwe iyo iri pamusoro yakanaka manhamba concordance yakanyorwa, chi-square bvunzo (MacNemar bvunzo) yakaratidza kuti mhedzisiro yeSansure Biotech assay yakave nemusiyano wakakosha uchienzaniswa neCRS (p <0.005). Kunyangwe iyo iri pamusoro yakanaka manhamba concordance yakanyorwa, chi-square bvunzo (MacNemar bvunzo) yakaratidza kuti mhedzisiro yeSansure Biotech assay yakave nemusiyano wakakosha uchienzaniswa neCRS (p <0.005). Несмотря на то, что было зафиксировано указанное выше хорошее статистическое соответствие, критерий хи-квадрат (критерий Макнезера, критерий Макнезей) Sansure Biotech имеет статистически значимое различие по сравнению с CRS (p <0,005). Kunyangwe chibvumirano chakanaka chenhamba pamusoro chakanyorwa, chi-square bvunzo (McNemar bvunzo) yakaratidza kuti mhedzisiro yeSansure Biotech assay yaive nemusiyano wakakosha uchienzaniswa neCRS (p <0.005).尽管记录了上述良好的统计一致性，但卡方检验（MacNemar 检验）表明，Sansure Biotech 检测的结果会相比具有统计学显着差异（p <0.005）.尽管 记录 上述 良好 统计 一致性 , 但 检验 （(macnemar 检验 表明 ， , sansure biotech 渎测具有 显着 （（p <0.005.。。。。。。。 Несмотря на отмеченное выше хорошее статистическое соответствие, критерий хи-квадрат (критерий Макнемара) показал статистическое соответствие, критерий хи-квадрат (критерий Макнемара) показал статистическое значим между анализом Sansure Biotech uye CRS. Pasinei nechibvumirano chakanaka chenhamba chataurwa pamusoro, bvunzo ye-chi-square (McNemar bvunzo) yakaratidza musiyano wakakosha (p <0.005) pakati peSansure Biotech assay neCRS.Mienzaniso mitanhatu (3.66%) yakawanikwa iine nhema dzisina kunaka kana ichienzaniswa neCRS (Supplementary Table 1); izvi zvakakosha zvikuru, kunyanya kupiwa simba rekuparadzira utachiona. Iyo data iri pamusoro inotsigirawo iyi yakaderera yekuona chiyero15.
Muchidzidzo ichi, Ct values ​​dzakatemerwa kune yega yega assay uye yakateerana chikuva, ine yakaderera zvinoreva Ct kukosha kwakataurwa muAbbott SARS-CoV-2 assay. Mhedzisiro iyi inogona kunge ine chekuita neAbbott's panguva imwe chete yakasanganiswa genetic test system yekuonekwa kweSARS-CoV-2. Naizvozvo, zvinoenderana neMufananidzo 1, 87.6% yeAbbott SARS-CoV-2 mhedzisiro yaive neCt kukosha pasi pe20. Ct tsika dziri pamusoro pe30 hadzina kunyorwa. Pamusoro pekushandisa kwaAbbott kweSARS-CoV-2 genetic test fomati, mhedzisiro iyi inogona kunge ine chekuita neiyo yakaderera yekuona muganho (32.5 RNA makopi/mL) 18, iyo yakadzikira katatu pane yakaderera muganho wekambani wezana makopi eRNA. /mL. ml)19.
Chidzidzo ichi chine zvimwe zvatisingakwanisi: chekutanga, hatina mwero/mareferensi nzira [sekuwanda kwehutachiwana kana kumwe ongororo yemurabhoritari (LDA)] nekuda kwekushaikwa kwezvekushandisa. Chechipiri, zvienzaniso zvose zvakashandiswa muchidzidzo ichi zvaive nasopharyngeal swabs, asi zvigumisiro zvakanga zvisingashandisi kune mamwe marudzi emuenzaniso, uye chechitatu, saizi yedu yemuenzaniso yakanga iri duku.
Ichi chidzidzo chakafananidza mashandiro echina rRT-PCR assays eSARS-CoV-2 achishandisa nasopharyngeal samples. Ese maassay ekuona aive nekuita kwakada kufanana, kunze kweiyo Sansure Biotech assay. Kunze kwezvo, iyo yakaderera positivity mwero wakaonekwa muSansure Biotech assay ichienzaniswa neCRS (p <0.05). Kunze kwezvo, iyo yakaderera positivity mwero wakaonekwa muSansure Biotech assay ichienzaniswa neCRS (p <0.05). Кроме того, в тесте Sansure Biotech был выявлен низкий процент положительных результатов по сравнению с CRS (p <0,05). Pamusoro pezvo, iyo Sansure Biotech bvunzo yakaratidza yakaderera muzana yemhedzisiro yakanaka kana ichienzaniswa neCRS (p <0.05).此外，与CRS 相比，Sansure Biotech 检测的阳性率较低(p <0.05).此外，与CRS 相比，Sansure Biotech 检测的阳性率较低(p <0.05). Кроме того, анализ Sansure Biotech имел более низкий уровень положительных результатов по сравнению с CRS (p <0,05). Uye zvakare, iyo Sansure Biotech assay yaive yakaderera positivity mwero uchienzaniswa neCRS (p <0.05).Iyo Sansure Biotech nCoV-2019 (RUO) ongororo yePPA, NPA uye chibvumirano chese chakapfuura 93.5% neCohen Kappa simba rekubvumirana kukosha kwe0.925. Chekupedzisira, iyo Sansure Biotech Assay (RUO) inoda kumwe kusimbiswa kuti ishandiswe muEthiopiya, uye kumwe kutsvagisa kunofanirwa kutariswa kuongorora zvichemo kubva kune vagadziri vega.
Kuenzanisa magadzirirwo ezvidzidzo akaitwa panzvimbo ina dzehutano muAddis Ababa, Eka Kotebe Hospital, Millennium Church Treatment Centre, Zewooditu Memorial Hospital, uye St. Peter Tuberculosis Specialist Hospital. Iyo data yakaunganidzwa pakati paZvita 1 na31, 2020. Zvivako zvekurapa zvechidzidzo ichi zvakasarudzwa nemaune zvichienderana nenhamba yadzo yepamusoro uye kuwanikwa kwenzvimbo huru dzekurapa muguta. Saizvozvo, zviridzwa, zvinosanganisira ABI 7500 uye Abbott m2000 chaiyo-nguva PCR zviridzwa, zvakasarudzwa maererano nekurudziro yevagadziri veNAAT reagent, uye mana PCR ekuona kits akasarudzwa pachidzidzo ichi, sezvo mazhinji marabhoritari muEthiopia akashandisa kanenge. vana vavo. Gene bvunzo, Abbott SARS-CoV-2 bvunzo, Sansure Biotech bvunzo, uye SARS-CoV-2 BGI bvunzo yakaitwa panguva yechidzidzo).
Kuyedzwa kweSARS-CoV-2 kwakaitwa kubva musi wa1 kusvika 30 Zvita 2020 pachishandiswa 3 ml yeViral Transport Medium (VTM) (Miraclean Technology, Shenzhen, China) kubva kuvanhu vari kuferefetwa nezveCCIDID-19 vanonzi EPHI. Nasopharyngeal samples dzakaunganidzwa nevateresi vakadzidziswa uye vakatumirwa kuEPHI mumapaketi matatu. Pamberi peiyo nucleic acid yekuparadzaniswa, imwe neimwe sampu inopihwa yakasarudzika nhamba yekuzivikanwa. Kudonhedza kunoitwa kubva kune yega sampuli pakarepo pakusvika uchishandisa manyore uye otomatiki nzira dzekubvisa. Nokudaro, nokuda kwekutsvaga kwekutsvaga kweAbbott m2000, 1.3 ml (kusanganisira 0.8 ml yakafa vhoriyamu uye 0.5 ml yekuwedzera inlet volume) yemuenzaniso yakatorwa kubva kune imwe neimwe sampuli uye yakapfuura neAbbott DNA Sample Preparation System (Abbott Molecular Inc. des Plaines, IL, USA). ) Batch ye96 [92 samples, maviri ekuona ma controls uye maviri asiri-template controls (NTC)] akasanganisirwa mukuita kwese (kudzosa nekuonekwa) kwemaround maviri eSARS-CoV-2 (EUA) munguva chaiyo. kuchera. Saizvozvo, kune zvinyorwa zvemanyorero, shandisa iwo masampula mamwechete (yeotomatiki kudhirowa uye kuwanikwa). Nekudaro, mukati memaitiro ese, 140 µl samples dzakatorwa uye dzakatorwa pachishandiswa QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Germany) mumabhechi e24 (kusanganisira 20 samples, maviri assay control uye maviri NTCs) pamusoro mapfumbamwe marounds. Manually eruates akakwenenzverwa uye akaonekwa achishandisa ABI 7500 thermal cycler uchishandisa SARS-CoV-2 BGI assay, Daan Gene assay, uye Sansure Biotech assay.
Otomatiki yekuzviparadzanisa nevamwe uye kucheneswa kweSARS-CoV-2 hutachiona RNA inoteedzera iyo magineti bead musimboti uchishandisa Abbott DNA sampuro yekugadzirira reagents. Kusashanda kwemasampuli uye solubilization yehutachiona hwehutachiona hunoitwa uchishandisa detergent ine guanidine isothiocyanate kuti denature protein uye inactivate RNase. Iyo RNA inobva yaparadzaniswa neprotein nechikamu chakasimba chekuparadzanisa kushandisa silica, kureva kuti guanidinium munyu uye alkaline pH ye lysis buffer inokurudzira kusunga kwe nucleic acids kune silica (SiO2). Nhanho yekusuka inobvisa mapuroteni akasara nemarara kuti abudise mhinduro yakajeka. Transparent RNA yakaparadzaniswa kubva kune silica-based microparticles uchishandisa chiridzwa chiridzwa magineti20,21. Kune rimwe divi, kuzvipatsanura kwemanyorero uye kucheneswa kweRNA kunoitwa neiyo spin column nzira uchishandisa centrifugation panzvimbo yemagineti kumira uye kupatsanurwa kwe microparticles kubva kune eluent.
Iyo Abbott Chaiyo-Nguva SARS-CoV-2 Detection Test (Abbott Molecular, Inc.) yakaitwa maererano nemirairo yemugadziri, iyo yakagamuchira EUA19,22 kubva kuWHO uye FDA. Mune iyi protocol, kusashanda kwemuenzaniso kusati kwatorwa kwakaitwa mumvura yekugezera pa56 ° C ye30 min. Mushure mekudzivirira kwehutachiona, nucleic acid yakaitwa paAbbott m2000 SP chiridzwa kubva ku0.5 ml VTM uchishandisa Abbott m2000 DNA sampuli yekugadzirira system. maererano nemugadziri. Kukwidziridzwa uye kucherechedzwa kwakaitwa pachishandiswa Abbott m2000 RT-PCR chiridzwa, uye kucherechedzwa kwemaviri kwakaitwa kuRdRp neN majini. ROX) uye VIC P (proprietary dye) yekutarisa uye kuona kwekutonga kwemukati, zvichibvumira kuonekwa panguva imwe chete yezvose zviri zviviri amplification zvigadzirwa 19.
Iyo yekukudza nzira yekuona yekiti iyi yakavakirwa pane imwe-nhanho RT-PCR tekinoroji. Iyo ORF1a/b neN majini akasarudzwa senzvimbo dzakachengetwa neDaan Gene Technology kuti ione kukwidziridzwa kwedunhu. Maprimers chaiwo uye fluorescent probes (N gene probes akanyorwa neFAM, ORF1a/b probes akanyorwa neVIC) akagadzirwa kuti aone SARS-CoV-2 RNA mumasample. Ekupedzisira eluent uye master musanganiswa akagadzirwa nekuwedzera 5 µl ye eluent kune 20 µl yemusanganiswa mukuru kune yekupedzisira vhoriyamu ye25 µl. Amplification uye kuonekwa kwakaitwa panguva imwe chete pane ABI 750024 chaiyo-nguva PCR chiridzwa.
Iwo ORF1a/b neN majini akaonekwa pachishandiswa Sansure Biotech nCoV-2019 Nucleic Acid Diagnostic Kit (fluorescent PCR kuonekwa). Gadzirira maprobe chaiwo ejini rega rega rinotarirwa nekusarudza chiteshi cheFAM chedunhu reORF1a/b uye ROX chiteshi cheN gene. Kune iyi kit yeassay, eluent uye master mix reagents inowedzerwa sezvizvi: gadzirira makumi matatu µl master mix reagent uye 20 µl eluted sampuro yekuonekwa/kukudza. Real-time PCR ABI 750025 yakashandiswa kukwidziridza/kuona.
Iyo SARS-CoV-2 BGI bvunzo ndeye fluorescent chaiyo-nguva rRT-PCR kit yekuongororwa kweCCIDID-19. Nzvimbo inotariswa iri munharaunda yeORF1a/b yeSARS-CoV-2 genome, inova nzira yekuona gene. Pamusoro pezvo, gene rekuchengetedza imba yemunhu β-actin igeni rakadzorwa mukati. Iyo master musanganiswa inogadzirwa nekusanganisa 20 µl ye master mix reagent uye 10 µl yeiyo yakabviswa RNA sampuro mugomba26. Iyo ABI 7500 fluorescent quantitative chaiyo-nguva PCR chiridzwa chakashandiswa kukwidziridza uye kuona. Yese nucleic acid amplification, PCR inomhanyisa mamiriro eyese assay, uye dudziro yemhedzisiro yakaitwa zvinoenderana neyakasiyana mirairo yemugadziri (Tafura 3).
Mukuongorora uku kwekuenzanisa, isu hatina kushandisa nzira yereferensi yekutarisa chibvumirano chezana (chakanaka, chisina kunaka, uye chakazara) uye mamwe maparamita ekuenzanisa kune ina ongororo. Kuenzanisa kwega kwega kwebvunzo kwakaitwa neCRS, muchidzidzo ichi CRS yakaiswa nemutemo wekuti "chero chakanaka" uye mhedzisiro yacho yakatemwa, kwete nebvunzo imwe chete, takashandisa zvingangoita zviviri zvakaenderana mhedzisiro. Pamusoro pezvo, munyaya yeCCIDID-19 kutapurirana, nhema dzisina kunaka mhedzisiro dzine njodzi pane mhedzisiro yenhema. Naizvozvo, kutaura "zvakanaka" nenzira kwayo sezvinobvira kubva pamugumisiro weCRS, bvunzo dzinokwana mbiri dzinofanira kunge dziri dzakanaka, zvichireva kuti mhedzisiro imwe chete yakanaka ingangobva kuEUA assay. Nekudaro, kubva pamabvunzo mana emhedzisiro, maviri kana anopfuura ebvunzo mhinduro inopa zvakafanana mhedzisiro inoonekwa seyakanaka kana negative18,27.
Dhata yakaunganidzwa pachishandiswa mafomu ekutora data akarongwa, kupinza data uye kuongororwa kwakaitwa pachishandiswa Excel statistical software uye SPSS vhezheni 23.0 yehuwandu hunotsanangura. Chibvumirano chakanaka, chakaipa, uye chese muzana chakaongororwa, uye chibodzwa cheKappa chakashandiswa kuona dhigirii rekubvumirana kwenzira yega yega neCRS. Hukoshi hweKappa hunodudzirwa sezvizvi: 0.01 kusvika 0.20 yechibvumirano chakapfava, 0.21 kusvika 0.40 chibvumirano chakajairika, 0.41-0.60 chibvumirano chepakati, 0.61-0.80 chibvumirano chikuru uye 0.81-0.99 yechibvumirano chakakwana28.
Ethical clearance yakawanikwa kubva kuYunivhesiti yeAddis Ababa uye zvese zvechidzidzo ichi zvakabvumidzwa neEthiopian Public Health Institute's Scientific Ethics Review Board. Nhamba yereferensi yeEPHI Ethics License ndeye EPHI/IRB-279-2020. Nzira dzese dzakashandiswa zvinoenderana nekurudziro uye zvakapihwa neEthiopian National Comprehensive Guidelines yekurapwa kweCOVID-19. Pamusoro pezvo, chibvumirano chakanyorwa chine ruzivo chakawanikwa kubva kune vese vatori vechikamu chidzidzo vasati vatora chikamu muchidzidzo.
Yese data yakawanikwa kana kuongororwa muchidzidzo ichi inosanganisirwa mune ino yakabudiswa chinyorwa. Data inotsigira zvakabuda muchidzidzo ichi inowanika kubva kumunyori kana zvakumbirwa.
Sangano Rinoona nezve Utano Pasi pose. Kurudziro dzeMarabhoritari Ekuyedza Strategies eCOVID-19: Yenguva Yekutungamira, Kurume 21, 2020 No. WHO/2019-nCoV/lab_testing/2020.1 (WHO, 2020).
Mouliou, DS, Pantazopoulos, I. & Gourgoulianis, KI COVID-19 akangwara kuongororwa muDhipatimendi reEmergency: Zvese-muKudzidzira. Mouliou, DS, Pantazopoulos, I. & Gourgoulianis, KI COVID-19 akangwara kuongororwa muDhipatimendi reEmergency: Zvese-muKudzidzira.Muliou, DS, Pantazopoulos, I. uye Gurgulianis, KI Intelligent kuongororwa kweCCIDID-19 mudhipatimendi rekukurumidzira: zvese zviri mukuita.Muliou DS, Pantazopoulos I. uye Gurgulyanis KI Intelligent kuongororwa kweCCIDID-19 mumadhipatimendi ekukurumidzira: kuguma-kusvika-kumagumo kubatanidzwa mukuita. Nyanzvi Reverend Respire. mushonga. 3, 263–272 (2022).
Mitchell, SL & St George, K. Ongororo yeCOVID19 ID ZVINO EUA assay. Mitchell, SL & St George, K. Ongororo yeCOVID19 ID ZVINO EUA assay.Mitchell, SL naSt. George, K. Ongororo yeCOVID19 ID ZVINO EUA assay.Mitchell SL naSt. George K. Ongororo yeCOVID19 ID ZVINO EUA assay. J. Clinical. Utachiona. 128, 104429. https://doi.org/10.1016/j.jcv.2020.104429 (2020).
WHO. Murabhoritari yekuona chirwere chekoronavirus 2019 (COVID-19) mune inofungidzirwa chirwere chevanhu. https://www.who.int/publications/i/item/10665-331501 (yakasvika 15 Nyamavhuvhu 2020) (WHO, 2020).
Udugama, B. et al. COVID-19 Kuongororwa: Zvirwere uye Kuedza Maturusi. ACS Nano 14(4), 3822–3835 (2020).
Syed S. et al. Kugadzwa kweKoreji yePathologists yekuEastern, Central uye Southern Africa - Dunhu Chikoro chePathology cheMiddle East neSouth Africa. Africa. J. Lab. mushonga. 9(1), 1-8 (2020).
Ethiopian Institute of Public Health, Federal Ministry of Health. Interim National Strategy uye Guidance for Laboratory Diagnostics yeCOVID-19. https://ephi.gov.et/images/novel_coronavirus/EPHI_PHEOC_COVID-19_Laboratory_Diagnosis_Eng.pdf (yakasvika 12 Nyamavhuvhu 2020) (EPHI, 2020).
Woloshin, S., Patel, N. & Kesselheim, AS Nhema dzisina kunaka bvunzo dzeSARS-CoV-2 matambudziko ehutachiona uye zvazvinoreva. Woloshin, S., Patel, N. & Kesselheim, AS Nhema dzisina kunaka bvunzo dzeSARS-CoV-2 matambudziko ehutachiona uye zvazvinoreva.Voloshin S., Patel N. uye Kesselheim AS Nhema-negative bvunzo dzeSARS-CoV-2 utachiona nemhedzisiro yazvo.Voloshin S., Patel N. uye Kesselheim AS Nhema-negative bvunzo dzekudenha uye kukanganisa kwehutachiona hweSARS-CoV-2. N. eng. J. Mushonga. 383(6), e38 (2020).
Mouliou, DS & Gourgoulianis, KI Nhema-yakanaka uye yenhema-yakaipa COVID-19 kesi: Kudzivirirwa kwekufema uye manejimendi ekugadzirisa, kubaya, uye mamwe maonero. Mouliou, DS & Gourgoulianis, KI Nhema-yakanaka uye yenhema-yakaipa COVID-19 kesi: Kudzivirirwa kwekufema uye manejimendi ekugadzirisa, kubaya, uye mamwe maonero. Mouliou, DS & Gourgoulianis, KI Ложноположительные uye ложноотрицательные случаи COVID-19: респираторная профилактика и сратегии лечения, ваийцишина перспективы. Mouliou, DS & Gourgoulianis, KI Nhema dzakanaka uye dzenhema dzakaipa nyaya dzeCCIDID-19: kudzivirira kufema uye nzira dzekurapa, kubaya uye nzira yekumberi.Muliu, DS naGurgulianis, KI Mhosva-yakanaka uye yenhema-yakashata nyaya dzeCCIDID-19: mazano ekudzivirira kufema uye kurapwa, kubaya uye nzira yekumberi. Nyanzvi Reverend Respire. mushonga. 15(8), 993–1002 (2021).
Mouliou, DS, Ioannis, P. & Konstantinos, G. COVID-19 kuongororwa mudhipatimendi rekukurumidzira: Kuona muti asi uchirasikirwa nesango. Mouliou, DS, Ioannis, P. & Konstantinos, G. COVID-19 kuongororwa mudhipatimendi rekukurumidzira: Kuona muti asi uchirasikirwa nesango.Mouliou, DS, Ioannis, P. naKonstantinos, G. COVID-19 Kuongororwa muDhipatimendi reEmergency: Ona Muti, Rasa Sango.Muliou DS, Ioannis P., naKonstantinos G. COVID-19 Kuongororwa Mudzimba dzeEmergency: Haana Sango Rakakwana reMiti. Kuonekwa. mushonga. J. https://doi.org/10.1136/emermed-2021-212219 (2022).
Degli-Angeli, E. et al. Kusimbisa uye Kusimbiswa kweKuongorora uye Kliniki Kuita kweAbbott RealTime SARS-CoV-2 Assay. J. Clinical. Utachiona. 129, 104474. https://doi.org/10.1016/j.jcv.2020.104474 (2020).
Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Kuenzanisa mashanu ekutanga seti kubva kwakasiyana genome dunhu reCOVID-19 yekuona kutapukirwa nehutachiona neyakajairwa RT-PCR. Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Kuenzaniswa kwemaseti mashanu ekutanga kubva munzvimbo dzakasiyana dzegenome dzeCCIDID-19 yekuona hutachiona hwehutachiona neyakajairwa RT-PCR.Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. uye Aflatunyan, B. Kuenzaniswa kwemaseti mashanu ekutanga kubva kumatunhu akasiyana eCOVID-19 genome yekuona hutachiona hwehutachiona neyakajairwa RT-PCR. Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. 比较來自COVID-19 不同基因组区域的五个引物组，用亞引物组检测病毒感染。 Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Kuenzanisa kwe5 genetics dzakasiyana dzeCOVID-19 yekuona hutachiona hwehutachiona neyakajairwa RT-PCR.Mollaei HR, Afshar AA, Kalantar-Neyestanaki D, Fazlalipour M. uye Aflatunyan B. Kuenzaniswa kwemaseti mashanu ekutanga kubva kumatunhu akasiyana eCOVID-19 genome yekuona hutachiona hwehutachiona neyakajairwa RT-PCR.Iran. J. Microbiology. 12(3), 185 (2020).
Goertzer, I. et al. Ekutanga mhedzisiro yenyika yekunze yemhando yekuongorora chirongwa chekuonekwa kweSARS-CoV-2 genome sequences. J. Clinical. Utachiona. 129, 104537. https://doi.org/10.1016/j.jcv.2020.104537 (2020).
Wang, M. et al. Ongororo Yekuongorora Kushanda KwemaKitsi mashanu eRT-PCR kune Yakanyanya Acute Respiratory Syndrome Coronavirus 2. J. Clinical. rabhoritari. anus. 35(1), e23643 (2021).
Wang B. et al. Ongororo yeanomwe anowanikwa mune zvekutengesa SARS-CoV-2 RNA yekuona kits muChina zvichibva pane chaiyo-nguva polymerase chain reaction (PCR). kliniki. Chemical. rabhoritari. mushonga. 58(9), e149–e153 (2020).
van Casteren, PB nevamwe. Kuenzanisa kweanomwe ekutengesa RT-PCR COVID-19 diagnostic kits. J. Clinical. Utachiona. 128, 104412 (2020).
Lu, Yu, nevamwe. Kuenzanisa kwekuita kwekuongorora kweviri PCR kits yekuonekwa kweSARS-CoV-2 nucleic acids. J. Clinical. rabhoritari. anus. 34(10), e23554 (2020).
Lefart, PR, etc. Ongororo yekuenzanisa yechina SARS-CoV-2 nucleic acid amplification test (NAAT) mapuratifomu yakaratidza kuti ID NOW kuita kwakaderedzwa zvakanyanya zvichienderana nemurwere uye mhando yemuenzaniso. kuongororwa. microbiology. Infect. diss. 99(1), 115200 (2021).
Abbott molecule. Abbott chaiyo-nguva SARS-CoV-2 yekuongorora pasuru yekuisa. https://www.molecular.abbott/us/en/products/infectious-disease/RealTime-SARS-CoV-2-Assay. 1-12. (Kubva Nyamavhuvhu 10, 2020) (2020).
Klein, S. et al. SARS-CoV-2 RNA yekuzviparadzanisa nekushandisa magineti mabhedhi ekukurumidza kutariswa hukuru-hukuru neRT-qPCR uye RT-LAMP. Hutachiona 12(8), 863 (2020).