Kuchita kwa mayeso anayi a nucleic acid amplification kuti azindikire SARS-CoV-2 ku Ethiopia

Zikomo pochezera Nature.com. Mukugwiritsa ntchito msakatuli wokhala ndi chithandizo chochepa cha CSS. Kuti mudziwe zambiri, tikupangira kuti mugwiritse ntchito msakatuli wosinthidwa (kapena kuletsa Compatibility Mode mu Internet Explorer). Kuphatikiza apo, kuti tiwonetsetse chithandizo chopitilira, tikuwonetsa tsambalo popanda masitayilo ndi JavaScript.
Imawonetsa carousel ya masilayidi atatu nthawi imodzi. Gwiritsani ntchito mabatani Akale ndi Otsatira kuti mudutse ma slide atatu nthawi imodzi, kapena gwiritsani ntchito mabatani otsetsereka kumapeto kuti mudutse ma slide atatu nthawi imodzi.
Chiyambireni matenda a coronavirus (COVID-19) a 2019, mayeso ambiri azamalonda a nucleic acid amplification (NAATs) apangidwa padziko lonse lapansi ndipo akhala oyesa wamba. Ngakhale kuti mayesero angapo anapangidwa mwamsanga ndikugwiritsidwa ntchito ku mayesero a ma laboratory, machitidwe a mayeserowa sanayesedwe m'malo osiyanasiyana. Chifukwa chake, kafukufukuyu anali ndi cholinga chowunika momwe ma assays a Abbott SARS-CoV-2, Daan Gene, BGI, ndi Sansure Biotech amagwiritsa ntchito Composite Reference Standard (CRS). Phunziroli linachitidwa ku Ethiopian Public Health Institute (EPHI) kuyambira 1 mpaka 30 December 2020. Zitsanzo za 164 za nasopharyngeal zinatulutsidwa pogwiritsa ntchito QIAamp RNA mini kit ndi dongosolo la kukonzekera chitsanzo cha Abbott DNA. Mwa zitsanzo 164, 59.1% zinali zabwino ndipo 40.9% zinali zoipa kwa CRS. Sansure Biotech positivity inali yotsika kwambiri poyerekeza ndi CRS (p <0.05). Sansure Biotech positivity inali yotsika kwambiri poyerekeza ndi CRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p <0,05). Zotsatira zabwino za Sansure Biotech zinali zotsika kwambiri poyerekeza ndi CRS (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 inali ndi zotsatira zabwino zochepa poyerekeza ndi CRS (p <0.05).Chigwirizano chonse cha kusanthula anayi chinali 96.3-100% poyerekeza ndi CRS. Kuphatikiza pa kutsika kwabwino kwa kuyesa kwa Sansure Biotech, magwiridwe antchito anayi anali pafupifupi ofanana. Chifukwa chake, kuyesa kwa Sansure Biotech [Research Only (RUO)] kumafunikira kutsimikizira kowonjezereka kuti kugwiritsidwe ntchito ku Ethiopia. Pomaliza, kafukufuku wowonjezera ayenera kuganiziridwa kuti ayese zoyeserera ndi zonena zoyenera za opanga.
Kuyesa kwa labotale ndi gawo la World Health Organisation (WHO) Strategic Plan for Coronavirus Disease 2019 (COVID-19) Preparedness and Response (SPRP). WHO ikulangiza kuti mayiko akuyenera kupanga ma laboratories kuti athe kukonzekera bwino, kuyang'anira bwino milandu, kukhala tcheru komanso kuyankha mofulumira pazovuta za umoyo wa anthu. Izi zikuwonetsa kuti ntchito ya labotale ndiyofunikira kwambiri pakuzindikiritsa matenda ndi miliri ya tizilombo toyambitsa matenda omwe akubwera ndikuwongolera kufalikira kwawo.
Kuzindikiridwa kwa COVID-19 kumafuna chidziwitso cha miliri ndi zamankhwala, zizindikiro zamunthu/zizindikiro, ndi data ya radiographic ndi labotale2. Chiyambireni mliri wa COVID-19 ku Wuhan, China, mayeso ambiri amalonda a nucleic acid amplification (NAATs) apangidwa padziko lonse lapansi. Real-time reverse transcript polymerase chain reaction (rRT-PCR) yakhala ikugwiritsidwa ntchito ngati njira yokhazikika komanso yodziwika bwino yodziwira matenda aacute kupuma kwa matenda amtundu wa 2 (SARS-CoV-2)3. Kuzindikira kwa mamolekyulu a SARS-CoV-2 nthawi zambiri kumatengera jini ya N (nucleocapsid protein), E (envelopu jini ya protein), ndi RdRp (RNA-dependent RNA polymerase gene) mu ORF1a/b (chithunzi chotseguka 1a/b) . gene) chigawo chodziwika kuchokera ku ma virus genome. Iwo amaonedwa ngati zigawo zazikulu zotetezedwa zomwe zimapezeka mu ma virus genomes a virus recognition4. Pakati pa majini awa, ma jini a RdRp ndi E ali ndi chidwi chodziwikiratu, pomwe jini ya N ili ndi chidwi chochepa chowunikira5.
Mayesero a PCR amatha kusiyanasiyana malinga ndi zinthu zosiyanasiyana monga: ma reagents otulutsa, ma amplification / detective reagents, njira yotulutsira, mtundu wa makina a PCR ndi zida zina. Pofika Epulo 2020, zida zopitilira 48 zowunikira matenda zochokera kumayiko asanu ndi anayi zalandila Emergency Use Authorization (EUA) pakuwunika kwa COVID-196. Ku Ethiopia, nsanja zopitilira 14 za PCR zenizeni zimagwiritsidwa ntchito pozindikira PCR ya SARS-CoV-2 m'mabungwe 26 aboma, kuphatikiza ABI 7500, Abbott m2000, Roche 48000 ndi Quant-studio7. Kuphatikiza apo, zida zoyesera zosiyanasiyana za PCR zilipo, monga kuyesa kwa Daan Gene, kuyesa kwa Abbott SARS-CoV-2, kuyesa kwa Sansure Biotech, ndi kuyesa kwa SARS-CoV-2 BGI. Ngakhale rRT-PCR imakhudzidwa kwambiri, odwala ena omwe ali ndi COVID-19 amafotokoza zabodza chifukwa chosowa makope a viral ribonucleic acid (RNA) m'zitsanzo chifukwa cha kusonkhanitsidwa molakwika, mayendedwe, kasungidwe ndi kasamalidwe, komanso kuyesa kwa labotale. mikhalidwe ndi zochita za ogwira ntchito8. Kuphatikiza apo, zitsanzo kapena kuwongolera kusamalidwa bwino, kukhazikika kwa cycle threshold (Ct), komanso kuyanjananso ndi ma nucleic acids ena kapena osagwira ntchito/otsalira a SARS-CoV-2 RNA kungayambitse zotsatira zabodza pamayesero a rRT-PCR9. Choncho, n'zoonekeratu kuti mayesero a PCR amatha kuzindikira zonyamulira za zidutswa za jini, chifukwa sangathe ngakhale kusiyanitsa pakati pa majeremusi ogwira ntchito, kotero kuti mayesero amatha kuzindikira onyamula osati odwala10. Chifukwa chake, ndikofunikira kuyesa magwiridwe antchito pogwiritsa ntchito njira zokhazikika pamakonzedwe athu. Ngakhale ma reagents ambiri a NAAT akupezeka ku Ethiopian Public Health Institute (EPHI) komanso m'dziko lonselo, palibe kuyerekeza koyerekeza kwawo komwe kwanenedwa. Chifukwa chake, kafukufukuyu anali ndi cholinga choyesa kufananiza kwa zida zomwe zikupezeka pamalonda kuti zizindikire SARS-CoV-2 ndi rRT-PCR pogwiritsa ntchito zitsanzo zachipatala.
Anthu 164 omwe akuwaganizira kuti ali ndi COVID-19 adaphatikizidwa mu kafukufukuyu. Zitsanzo zambiri zidachokera kuzipatala (118/164 = 72%), pomwe otsala 46 (28%) adachokera kumalo osapangira chithandizo. Mwa omwe sanalandire chithandizo pakatikati, 15 (9.1%) anali ndi milandu yomwe akuwakayikira ndipo 31 (18.9%) adalumikizana ndi milandu yotsimikizika. Otsatira makumi asanu ndi anayi mphambu atatu (56.7%) anali amuna, ndipo zaka (± SD) za omwe adatenga nawo gawo zinali zaka 31.10 (± 11.82).
Mu kafukufukuyu, ziwopsezo zabwino ndi zoyipa za mayeso anayi a COVID-19 zidatsimikiziridwa. Chifukwa chake, mitengo yabwino ya Abbott SARS-CoV-2 assay, Daan Gene 2019-nCoV assay, SARS-CoV-2 BGI assay, ndi Sansure Biotech 2019-nCoV assay anali 59.1%, 58.5%, 57.9% ndi 55.5% motsatana. . Zolemba zabwino ndi zoipa (CRS) zowerengera zinali 97 (59.1%) ndi 67 (40.9%), motsatira (Table 1). Mu kafukufukuyu, tanthauzo la CRS lidatengera lamulo la "zabwino zilizonse", zomwe mwa zotsatira zinayi zoyeserera, ziwiri kapena zingapo zoyeserera zomwe zidapereka zomwezo zimawonedwa ngati zabwino kapena zoipa.
Mu kafukufukuyu, tapeza mgwirizano wolakwika (NPA) wa 100% (95% CI 94.6-100) pazowunikira zonse poyerekeza ndi CRS. Kusanthula kwa Sansure Biotechnology kunawonetsa PPA yochepa ya 93.8% (95% CI 87.2-97.1) ndi kusanthula kwa Daan Gene 2019-nCoV kunali ndi mgwirizano wonse wa 99.4% (95% CI 96.6-99.9). Mosiyana ndi izi, mgwirizano wonse pakati pa kuyesa kwa SARS-CoV-2 BGI ndi kuyesa kwa Sansure Biotech 2019-nCoV kunali 98.8% ndi 96.3%, motsatana (Table 2).
Cohen's kappa coefficient of coefficient of the CRS and Abbott SARS-CoV-2 assay coefficient anali ogwirizana kwathunthu (K = 1.00). Momwemonso, mayendedwe a Cohen's kappa omwe adadziwika ndi Daan Gene 2019-nCoV, SARS-CoV-2 BGI, ndi Sansure Biotech 2019-nCoV nawonso amagwirizana kwathunthu ndi CRS (K ≥ 0.925). Pakuyerekeza uku, mayeso a chi-square (mayeso a McNemar) adawonetsa kuti zotsatira zoyesa za Sansure Biotech 2019-nCoV zinali zosiyana kwambiri ndi zotsatira za CRS (p = 0.031) (Table 2).
Monga momwe tawonetsera mkuyu.1 peresenti ya mtengo wotsika kwambiri wa Ct (<20 Ct) wa Abbott SARS-CoV-2 assay (wophatikiza RdRp ndi N jini) anali 87.6% ndi ORF1a/b gene Ct mtengo wa Sansure Biotech 2019-nCoV assay adawonetsa kuti kuchuluka kwa otsika Mtengo wa Ct (<20 Ct) unali 50.3% ndi mtengo wapamwamba wa Ct (36-40 Ct) anali 3.2%. 1 peresenti ya mtengo wotsika kwambiri wa Ct (<20 Ct) wa Abbott SARS-CoV-2 assay (wophatikiza RdRp ndi N jini) anali 87.6% ndi ORF1a/b gene Ct mtengo wa Sansure Biotech 2019-nCoV assay adawonetsa kuti kuchuluka kwa otsika Mtengo wa Ct (<20 Ct) unali 50.3% ndi mtengo wapamwamba wa Ct (36-40 Ct) anali 3.2%.Monga momwe tawonetsera mkuyu.5 2019-nCoV показало что процент низкого значения Ct (< 20 Ct) составлял 50,3%, а высокое значение Ct (36–40 Ct) составляло 3,2%. 1, peresenti ya mtengo wotsika kwambiri wa Ct (<20 Ct) kusanthula kwa Abbott SARS-CoV-2 (gene RdRp ndi N) inali 87.6%, ndipo mtengo wa Ct wa ORF1a/b kusanthula jini kwa Sansure Biotech 2019-nCoV kunawonetsa kuti peresenti ya mtengo wotsika wa Ct (<20 Ct) unali 50.3%, ndi mtengo wapamwamba wa Ct (36-40 Ct) adawerengera 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1sure19-V0tech 87.6%2,San 187.6%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. Monga momwe zasonyezedwera pa Chithunzi 1, peresenti yotsika kwambiri ya Ct (<20 Ct) ya mayeso a Abbott SARS-CoV-2 (kuphatikiza RdRp ndi N jini) ndi 87.6%, mtengo wa ORF1a/b Ct wa Sansure Biotech 2019-nCoV test ikuwonetsa Ct 值(<20 Ct) yotsika ndi 50.3%,高Ct 值(36–40 Ct) 的 peresenti ndi 3.2%. Как показано pa рисунке 1, анализ Abbott SARS-CoV-2 (сочетающий гены RdRp и N) имел самое низкое процентное значение Ct (< 20 Ct% , 8, 8, 7 гена ORF1a/b в исследовании Sansure Biotech 2019- Анализ nCoV показал низкий Ct. Monga tawonetsera pa Chithunzi 1, kuyesa kwa Abbott SARS-CoV-2 (kuphatikiza ma RdRp ndi N) kunali ndi Ct yotsika kwambiri (<20 Ct) pa 87.6%, pomwe mtengo wa Ct wa jini ya ORF1a/b mu Sansure. Kafukufuku wa Biotech 2019 - Kuwunika kwa nCoV kunawonetsa Ct yotsika. Процент значений (< 20 Ct) составил 50,3%, а процент высоких значений Ct (36–40 Ct) составил 3,2%. Maperesenti (< 20 Ct) anali 50.3%, ndipo kuchuluka kwa ma Ct apamwamba (36-40 Ct) anali 3.2%.Mayeso a Abbott SARS-CoV-2 B adalemba ma Ct apamwamba kuposa 30. Komano, pa BGI SARS-CoV-2 assay ORF1a / b jini anali mkulu Ct mtengo (> 36 Ct) peresenti anali 4% (mkuyu. 1). Komano, pa BGI SARS-CoV-2 assay ORF1a / b jini anali mkulu Ct mtengo (> 36 Ct) peresenti anali 4% (mkuyu. 1). С другой стороны, в анализе BGI SARS-CoV-2 gulu ORF1a/b ndi высокое значение Ct (> 36 Ct), процент которого составлял 4% (mzere 1). Kumbali ina, pakuwunika kwa jini ya BGI SARS-CoV-2 ORF1a/b inali ndi mtengo wapamwamba wa Ct (> 36 Ct), kuchuluka kwake komwe kunali 4% (mkuyu 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）. Kumbali ina, pakuzindikira kwa BGI SARS-CoV-2, kuchuluka kwa jini ya ORF1a/b yokhala ndi mtengo wapamwamba wa Ct (>36 Ct) ndi 4% (Chithunzi 1). С другой стороны, в анализе BGI SARS-CoV-2 процент генов ORF1a/b с высокими значениями Ct (>36 Ct) составил 4% (рис. 1). Kumbali ina, pakuwunika kwa BGI SARS-CoV-2, kuchuluka kwa majini a ORF1a/b okhala ndi ma Ct apamwamba kwambiri (> 36 Ct) anali 4% (mkuyu 1).
Mu phunziro ili, tinatenga zitsanzo za nasopharyngeal 164. Pa mitundu yonse ya zoyesa, kudzipatula kwa RNA ndi kukulitsa kunachitika pogwiritsa ntchito njira ndi zida zomwe amalangizidwa ndi opanga.
Kafukufukuyu adawonetsa kuti mayeso a Abbott a SARS-CoV-2 ali ndi magwiridwe antchito ofanana ndi a CRS, okhala ndi 100% zabwino, zoyipa, komanso concordance yonse. Mgwirizano wa Cohen wa kappa ndi 1.00, zomwe zikuwonetsa kuvomerezana kwathunthu ndi CRS. Kafukufuku wofananira ndi University of Washington ku US adapeza kuti kukhudzika konse komanso kutsimikizika kwa mayeso a Abbott a SARS-CoV-2 anali 93% ndi 100%, motsatana, poyerekeza ndi labotale-determined assay (LDA) ya CDC. . 11. Njira yodziwira ya Abbott SARS-CoV-2 imachokera ku kuzindikira kophatikizana kophatikizana kwa jini ya N ndi RdRp, popeza majini onse amakhudzidwa kwambiri, kuchepetsa zolakwika zabodza12. Kafukufuku yemwe adachitika ku Vienna, Austria adawonetsanso kuti ma voliyumu akulu ochotsa ndikuzindikira ma voliyumu amachepetsa kuchepetsedwa ndikuwonjezera kuzindikira13. Chifukwa chake, kufanana kwabwino kwa Abbott pakuyesa kwa SARS-CoV-2 kumatha kulumikizidwa ndi njira yodziwira nsanja yomwe nthawi yomweyo imazindikira majini ophatikizika, imatulutsa zitsanzo zambiri (0.5 ml), ndikugwiritsa ntchito zochulukirapo (40 µl).
Zotsatira zathu zinawonetsanso kuti kudziwika kwa kuyesa kwa majini a Daan kunali kofanana ndi kwa CRS. Izi zikugwirizana ndi kafukufuku14 yemwe adachitika ku Yunivesite ya Anhui ku Huainan, China, ndi zomwe wopanga ananena kuti 100% agwirizana bwino. Ngakhale malipoti a zotsatira zosasinthika, chitsanzo chimodzi chinali cholakwika pambuyo poyesanso zomwezo, koma zinali zabwino pamayesero a Abbott SARS-CoV-2 ndi Sansure Biotech nCoV-2019. Izi zikutanthawuza kuti pakhoza kukhala kusiyana kwa zotsatira pamitundu yosiyanasiyana ya mayesero. Komabe, mu kafukufuku wopangidwa ku China15, zotsatira za kuyesa kwa Daan Gene kunali kosiyana kwambiri (p <0.05) poyerekeza ndi kuwunika kwawo komwe kumatanthauzidwa ndi labu. Komabe, mu kafukufuku wopangidwa ku China15, zotsatira za kuyesa kwa Daan Gene kunali kosiyana kwambiri (p <0.05) poyerekeza ndi kuwunika kwawo komwe kumatanthauzidwa ndi labu. Тем не менее, в исследовании, проведенном Китае15, результат анализа Daan Gene значительно отличался (p <0,05) от их лаборалинся от их лаборатор. Komabe, mu kafukufuku ku China15, zotsatira za kusanthula kwa Daan Gene zinali zosiyana kwambiri (p <0.05) kuchokera ku kafukufuku wawo wa labotale.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 Китае15, результаты генетического теста Daan значительно отличались (p <0,05) по сравнетического теста Daan тестом. Komabe, mu kafukufuku ku China15, zotsatira za mayeso a majini a Daan zinali zosiyana kwambiri (p <0.05) poyerekeza ndi mayeso ake a labotale.Kusiyanaku kungakhale chifukwa cha kukhudzika kwa mayeso owunikira kuti azindikire SARS-CoV-2, ndipo maphunziro owonjezera angakhale ofunikira kuti adziwe chomwe chimayambitsa.
Kuphatikiza apo, kafukufuku wathu adawunikira magwiridwe antchito oyerekeza a SARS-CoV-2 BGI assay ndi CRS, akuwonetsa mgwirizano wabwino kwambiri (PPA = 97.9%), mgwirizano wopanda pake (NPA = 100%), ndi mgwirizano wonse pakati pa amuna ndi akazi ( OPA). ). = 98.8%). Makhalidwe a Cohen a Kappa adawonetsa mgwirizano wabwino (K = 0.975). Maphunziro ku Netherlands16 ndi China15 awonetsa zotsatira zofananira. Kuyesa kwa SARS-CoV-2 BGI ndi kuyesa kwa jini imodzi (ORF1a/b) pogwiritsa ntchito 10 µl kukulitsa/kuzindikira. Ngakhale kuvomerezana kowerengera bwino ndi zotsatira zathu zowunikira, kuwunikaku kudaphonya zitsanzo ziwiri zabwino (1.22%) za zitsanzo zonse. Izi zitha kukhala ndi tanthauzo lalikulu lazachipatala pamayendedwe opatsirana kwa odwala komanso ammudzi.
Kusanthula kwina koyerekeza komwe kudaphatikizidwa mu kafukufukuyu kunali kuyesa kwa Sansure Biotech nCoV-2019 rRT-PCR (RUO); machesi onse anali 96.3%. Mphamvu ya mgwirizano idatsimikiziridwanso ndi mtengo wa Cohen's Kappa, womwe unali 0.925, kusonyeza mgwirizano wonse ndi CRS. Apanso, zotsatira zathu ndi zofanana ndi maphunziro omwe anachitika ku Central South University ku Changsha, China, komanso ku Clinical Laboratory Department ya Liuzhou People's Hospital, Liuzhou City, China17. Ngakhale ziwerengero zabwino zomwe zili pamwambazi zidajambulidwa, mayeso a chi-square (mayeso a MacNemar) adawonetsa kuti zotsatira za kuyesa kwa Sansure Biotech zakhala ndi kusiyana kwakukulu poyerekeza ndi CRS (p <0.005). Ngakhale ziwerengero zabwino zomwe zili pamwambazi zidajambulidwa, mayeso a chi-square (mayeso a MacNemar) adawonetsa kuti zotsatira za kuyesa kwa Sansure Biotech zakhala ndi kusiyana kwakukulu poyerekeza ndi CRS (p <0.005). Несмотря на то, что было зафиксировано указанное выше хорошее статистическое соответствие, критерий хи-квадрат (критерий Макнезей, критерий хи-квадрат) Sansure Biotech ndimeет статистически значимое различие по сравнению с CRS (p <0,005). Ngakhale mgwirizano wabwino wa ziwerengero pamwambapa udalembedwa, kuyesa kwa chi-square (kuyesa kwa McNemar) kunawonetsa kuti zotsatira za kuyesa kwa Sansure Biotech zinali ndi kusiyana kwakukulu poyerekeza ndi CRS (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 статистистически значи0,у между анализом Sansure Biotech ndi CRS. Ngakhale pali mgwirizano wabwino wa ziwerengero zomwe tazitchula pamwambapa, kuyesa kwa chi-square (kuyesa kwa McNemar) kunawonetsa kusiyana kwakukulu (p <0.005) pakati pa Sansure Biotech assay ndi CRS.Zitsanzo zisanu ndi chimodzi (3.66%) zinapezeka kuti ndizolakwika poyerekeza ndi CRS (Supplementary Table 1); izi ndizofunikira kwambiri, makamaka chifukwa cha kufalikira kwa kachilomboka. Zomwe zili pamwambazi zimathandiziranso kutsika kodziwika bwino15.
Mu kafukufukuyu, mayendedwe a Ct adatsimikiziridwa pachiyeso chilichonse ndi nsanja, ndi mtengo wotsika kwambiri wa Ct womwe udanenedwa mu mayeso a Abbott SARS-CoV-2. Zotsatirazi zitha kukhala zokhudzana ndi njira ya Abbott yophatikizira ma genetic kuyesa kwa SARS-CoV-2. Choncho, malinga ndi Chithunzi 1, 87.6% ya zotsatira za Abbott SARS-CoV-2 zinali ndi Ct values ​​​​pansi pa 20. Zotsatira zochepa chabe (12.4%) zinali mu 20-30. Ma Ct opitilira 30 sanalembedwe. Kuphatikiza pakugwiritsa ntchito kwa Abbott mawonekedwe oyesa ma genetic a SARS-CoV-2, zotsatirazi zitha kukhala zokhudzana ndi malire otsika (makopi 32.5 RNA / mL) 18, omwe ndi otsika katatu kuposa malire otsika amakampani a makope 100 a RNA. /ml. ml) 19.
Kafukufukuyu ali ndi malire ena: choyamba, tilibe njira zodziwika bwino [monga kuchuluka kwa ma virus kapena kuyesa kwina kwa labotale (LDA)] chifukwa chosowa zinthu. Chachiwiri, zitsanzo zonse zomwe zimagwiritsidwa ntchito mu phunziroli zinali za nasopharyngeal swabs, pamene zotsatira zake sizinagwiritsidwe ntchito ku mitundu ina ya zitsanzo, ndipo chachitatu, kukula kwathu kwachitsanzo kunali kochepa.
Kafukufukuyu adafanizira magwiridwe antchito a ma rRT-PCR anayi a SARS-CoV-2 pogwiritsa ntchito zitsanzo za nasopharyngeal. Mayesero onse ozindikira anali ndi magwiridwe antchito ofanana, kupatula kuyesa kwa Sansure Biotech. Kupatula apo, kutsika kwabwino kudazindikirika pakuyesa kwa Sansure Biotech poyerekeza ndi CRS (p <0.05). Kupatula apo, kutsika kwabwino kudazindikirika pakuyesa kwa Sansure Biotech poyerekeza ndi CRS (p <0.05). Кроме того, в тесте Sansure Biotech был выявлен низкий процент положительных результатов по сравнению с CRS (p <0,05). Kuphatikiza apo, mayeso a Sansure Biotech adawonetsa zotsatira zotsika poyerekeza ndi CRS (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 ndimeл более низкий уровень положительных результатов по сравнению с CRS (p <0,05). Kuphatikiza apo, kuyesa kwa Sansure Biotech kunali ndi chiwongola dzanja chochepa poyerekeza ndi CRS (p <0.05).Kusanthula kwa Sansure Biotech nCoV-2019 (RUO) kwa PPA, NPA ndi mgwirizano wonse wadutsa 93.5% ndi mphamvu ya Cohen Kappa yogwirizana ndi 0.925. Pomaliza, Sansure Biotech Assay (RUO) ikufunika kutsimikiziridwa kwina kuti igwiritsidwe ntchito ku Ethiopia, ndipo kafukufuku wowonjezera akuyenera kuganiziridwa kuti awunike zonena za opanga pawokha.
Kapangidwe ka kafukufuku woyerekeza kunachitika m'zipatala zinayi ku Addis Ababa, Chipatala cha Eka Kotebe, Millennium Church Treatment Center, Zewooditu Memorial Hospital, ndi Chipatala cha St. Peter's Tuberculosis Specialist. Deta inasonkhanitsidwa pakati pa Disembala 1 ndi 31, 2020. Zipatala za kafukufukuyu zidasankhidwa mwadala kutengera kuchuluka kwa milandu komanso kupezeka kwa zipatala zazikulu mumzindawu. Momwemonso, zida, kuphatikiza zida za ABI 7500 ndi Abbott m2000 zenizeni zenizeni za PCR, zidasankhidwa malinga ndi malingaliro a opanga reagent a NAAT, ndipo zida zinayi zozindikira za PCR zidasankhidwa pa kafukufukuyu, monga ma laboratories ambiri ku Ethiopia adagwiritsa ntchito osachepera. anayi a iwo. Mayeso a Gene, mayeso a Abbott SARS-CoV-2, mayeso a Sansure Biotech, ndi mayeso a SARS-CoV-2 BGI omwe adachitika panthawi yophunzira).
Kuyesa kwa SARS-CoV-2 kunachitika kuyambira pa 1 mpaka 30 Disembala 2020 pogwiritsa ntchito 3 ml ya Viral Transport Medium (VTM) (Miraclean Technology, Shenzhen, China) kuchokera kwa anthu omwe akufufuzidwa chifukwa cha COVID-19 otchedwa EPHI. Zitsanzo za nasopharyngeal zidasonkhanitsidwa ndi osonkhanitsa ophunzitsidwa bwino ndikutumizidwa ku EPHI m'mapaketi atatu. Asanayambe kudzipatula kwa nucleic acid, chitsanzo chilichonse chimapatsidwa nambala yapadera. M'zigawo ikuchitika aliyense chitsanzo yomweyo pa kufika ntchito Buku ndi basi m'zigawo njira. Choncho, kwa basi m'zigawo za Abbott m2000, 1.3 ml (kuphatikiza 0,8 ml ya akufa voliyumu ndi 0,5 ml m'zigawo zolowera voliyumu) ​​wa chitsanzo anali yotengedwa chitsanzo aliyense ndi kudutsa Abbott DNA Zitsanzo Kukonzekera System (Abbott Molecular Inc. des Plaines, IL, USA). ) Gulu la 96 [zitsanzo 92, zowongolera ziwiri zodziwikiratu ndi zowongolera ziwiri zopanda template (NTC)] zidaphatikizidwa munjira yonse (kubweza ndi kuzindikira) ya mizere iwiri ya SARS-CoV-2 (EUA) munthawi yeniyeni. migodi. Momwemonso, pochotsa pamanja, gwiritsani ntchito zitsanzo zomwezo (zotulutsa zokha ndi kuzipeza). Chifukwa chake, munthawi yonseyi, zitsanzo za 140 µl zidatulutsidwa ndikuchotsedwa pogwiritsa ntchito QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Germany) m'magulu a 24 (kuphatikiza zitsanzo za 20, zowongolera ziwiri ndi ma NTC awiri) pamizere isanu ndi inayi. Zotulutsa pamanja zidakulitsidwa ndikuzindikiridwa pogwiritsa ntchito ABI 7500 thermal cycler pogwiritsa ntchito SARS-CoV-2 BGI assay, Daan Gene assay, ndi Sansure Biotech assay.
Kudzipatula ndi kuyeretsa kwa SARS-CoV-2 viral RNA kumatsatira mfundo ya maginito ya bead pogwiritsa ntchito ma reagents okonzekera a Abbott DNA. Inactivation wa zitsanzo ndi solubilization wa tizilombo particles ikuchitika ntchito detergent munali guanidine isothiocyanate kuti denature mapuloteni ndi inactivate RNase. RNA imasiyanitsidwa ndi mapuloteni ndi kupatukana kolimba kwa gawo pogwiritsa ntchito silika, mwachitsanzo, mchere wa guanidinium ndi alkaline pH wa lysis buffer zimalimbikitsa kumanga nucleic acid ku silika (SiO2). Njira yotsuka imachotsa mapuloteni otsala ndi zinyalala kuti apange yankho lomveka bwino. Transparent RNA imasiyanitsidwa ndi ma microparticles opangidwa ndi silika pogwiritsa ntchito maginito a chida20,21. Komano, kudzipatula pamanja ndi kuyeretsedwa kwa RNA kumayendetsedwa ndi njira yozungulira ndime yogwiritsa ntchito centrifugation m'malo mwa maginito ndikulekanitsa ma microparticles kuchokera ku eluent.
Mayeso a Abbott Real-Time SARS-CoV-2 Detection Test (Abbott Molecular, Inc.) adachitidwa molingana ndi malangizo a wopanga, omwe adalandira EUA19,22 kuchokera ku WHO ndi FDA. Mu protocol iyi, kusagwira ntchito kwachitsanzo kusanachitike kunachitika mu osamba amadzi pa 56 ° C kwa mphindi 30. Pambuyo poyambitsa kachilomboka, kutulutsa kwa nucleic acid kunachitika pa chida cha Abbott m2000 SP kuchokera ku 0.5 ml VTM pogwiritsa ntchito njira yokonzekera zitsanzo za Abbott m2000 DNA. malinga ndi wopanga. Kukulitsa ndi kuzindikira kunachitika pogwiritsa ntchito chida cha Abbott m2000 RT-PCR, ndipo kuzindikira kwapawiri kunachitika kwa majini a RdRp ndi N. ROX) ndi VIC P (utoto wa mwiniwake) pofuna kuwongolera ndi kuzindikira zowongolera zamkati, kulola kuzindikira munthawi yomweyo zinthu zonse zokulitsa 19.
Njira yodziwira kukulitsa kwa zida izi idakhazikitsidwa paukadaulo wa RT-PCR wagawo limodzi. Ma jini a ORF1a/b ndi N adasankhidwa ngati madera otetezedwa ndi Daan Gene Technology kuti azindikire kukula kwa chigawo. Zoyambira zenizeni ndi ma probes a fulorosenti (ma jini a N olembedwa ndi FAM, ORF1a/b olembedwa ndi VIC) adapangidwa kuti azitha kuzindikira SARS-CoV-2 RNA m'masamples. Zosakaniza zomaliza ndi zaluso zidakonzedwa powonjezera 5 µl ya eluent ku 20 µl ya master mix ku voliyumu yomaliza ya 25 µl. Kukulitsa ndi kuzindikira kunachitika nthawi imodzi pa chida cha PCR cha ABI 750024 chenicheni.
Mitundu ya ORF1a/b ndi N idapezedwa pogwiritsa ntchito Sansure Biotech nCoV-2019 Nucleic Acid Diagnostic Kit (kuzindikira kwa fulorosenti PCR). Konzani zofufuza zenizeni za jini iliyonse yomwe mukufuna kusankha posankha njira ya FAM ya dera la ORF1a/b ndi kanjira ka ROX ka jini ya N. Pa zida zoyesererazi, zosakaniza zosawoneka bwino ndi zosakaniza zimawonjezedwa motere: konzani 30 µl master mix reagent ndi 20 µl eluted zitsanzo kuti zizindikiridwe/kukulitsa. Real-time PCR ABI 750025 idagwiritsidwa ntchito pokulitsa / kuzindikira.
Mayeso a SARS-CoV-2 BGI ndi zida za fulorosenti zenizeni za rRT-PCR zozindikiritsa COVID-19. Dera lomwe mukufuna lili m'chigawo cha ORF1a/b cha SARS-CoV-2 genome, yomwe ndi njira yodziwira jini imodzi. Kuphatikiza apo, jini yosamalira m'nyumba ya anthu β-actin ndi jini yomwe imayendetsedwa mkati. Kusakaniza kwa master kumakonzedwa ndikusakaniza 20 µl ya master mix reagent ndi 10 µl ya RNA yotengedwa m'chitsime26. Chida cha ABI 7500 fulorosenti yochulukira nthawi yeniyeni ya PCR idagwiritsidwa ntchito pokulitsa ndi kuzindikira. Ma nucleic acid amplification onse, PCR imayendetsa mikhalidwe pa kuyesa kulikonse, ndipo kutanthauzira kwazotsatira kunachitika molingana ndi malangizo a wopanga (Table 3).
Pakuwunika kofananizaku, sitinagwiritse ntchito njira yofananira kuti tidziwe mgwirizano wa peresenti (zabwino, zoyipa, ndi zonse) ndi magawo ena ofananira pazowunikira zinayi. Kuyerekeza kulikonse kunachitika ndi CRS, mu kafukufukuyu CRS idakhazikitsidwa ndi lamulo la "zabwino zilizonse" ndipo zotsatira zake zidatsimikiziridwa, osati ndi mayeso amodzi, tidagwiritsa ntchito zotsatira zosachepera ziwiri zofananira. Kuphatikiza apo, pankhani yakufalitsa COVID-19, zotsatira zabodza ndizowopsa kuposa zotsatira zabodza. Chifukwa chake, kunena kuti "zabwino" molondola momwe ndingathere kuchokera pazotsatira za CRS, mayeso osachepera awiri ayenera kukhala abwino, kutanthauza kuti mwina chotsatira chimodzi chabwino chimachokera ku kuyesa kwa EUA. Chifukwa chake, mwa zotsatira zinayi zoyezetsa, ziwiri kapena zingapo zoyeserera zomwe zimapereka zotsatira zofanana zimawonedwa kuti zilidi zabwino kapena zoipa18,27.
Deta inasonkhanitsidwa pogwiritsa ntchito mafomu opangira deta, kulowa ndi kusanthula deta kunachitika pogwiritsa ntchito pulogalamu ya Excel statistical software ndi SPSS version 23.0 paziwerengero zofotokozera. Mgwirizano wabwino, woipa, ndi wathunthu unawunikidwa, ndipo mphambu ya Kappa idagwiritsidwa ntchito kudziwa kuchuluka kwa mgwirizano wa njira iliyonse ndi CRS. Makhalidwe a Kappa amatanthauziridwa motere: 0.01 mpaka 0.20 pa mgwirizano wochepa, 0.21 mpaka 0.40 pa mgwirizano wamba, 0.41-0.60 pa mgwirizano wapakati, 0.61-0.80 pa mgwirizano waukulu ndi 0.81-0.99 pa mgwirizano wonse28.
Chilolezo cha makhalidwe abwino chinapezedwa kuchokera ku yunivesite ya Addis Ababa ndipo ndondomeko zonse zoyesera za kafukufukuyu zinavomerezedwa ndi Bungwe la Scientific Ethics Review Board la Ethiopian Public Health Institute. Nambala yolozera ya EPHI Ethics License ndi EPHI/IRB-279-2020. Njira zonse zidagwiritsidwa ntchito molingana ndi malingaliro ndi zoperekedwa ndi Ethiopian National Comprehensive Guidelines for the Treatment of COVID-19. Kuonjezera apo, chilolezo cholembedwa cholembedwa chinapezedwa kuchokera kwa onse omwe adachita nawo kafukufuku asanatenge nawo mbali mu phunziroli.
Zonse zomwe zapezedwa kapena kufufuzidwa mu phunziroli zikuphatikizidwa m'nkhani yofalitsidwayi. Deta yochirikiza zotsatira za kafukufukuyu ikupezeka kuchokera kwa mlembi wake akafuna.
Bungwe Loyanganira Thanzi Padziko Lonse la Pansi. Malangizo a Njira Zoyezera Ma Laboratory a COVID-19: Interim Guidance, Marichi 21, 2020 No. WHO/2019-nCoV/lab_testing/2020.1 (WHO, 2020).
Mouliou, DS, Pantazopoulos, I. & Gourgoulianis, KI COVID-19 kuzindikira kwanzeru mu dipatimenti ya Emergency Department: All-in in Practice. Mouliou, DS, Pantazopoulos, I. & Gourgoulianis, KI COVID-19 kuzindikira kwanzeru mu dipatimenti ya Emergency Department: All-in in Practice.Muliou, DS, Pantazopoulos, I. ndi Gurgulianis, KI Intelligent matenda a COVID-19 mu dipatimenti yadzidzidzi: chilichonse chikuchitika.Muliou DS, Pantazopoulos I. ndi Gurgulyanis KI Intelligent matenda a COVID-19 m'madipatimenti azadzidzidzi: kuphatikiza-kumaliza-kuphatikizana muzochita. Katswiri Reverend Respire. mankhwala. 3, 263–272 (2022).
Mitchell, SL & St George, K. Kuwunika kwa COVID19 ID TSOPANO EUA assay. Mitchell, SL & St George, K. Kuwunika kwa COVID19 ID TSOPANO EUA assay.Mitchell, SL ndi St. George, K. Kuwunika kwa COVID19 ID TSOPANO EUA assay.Mitchell SL ndi St. George K. Kuwunika kwa COVID19 ID TSOPANO EUA assay. J. Zachipatala. Kachilombo. 128, 104429. https://doi.org/10.1016/j.jcv.2020.104429 (2020).
WHO. Kuzindikira kwa labotale kwa matenda a coronavirus 2019 (COVID-19) m'matenda omwe akuganiziridwa kuti ndi anthu. https://www.who.int/publications/i/item/10665-331501 (yofikira pa 15 Ogasiti 2020) (WHO, 2020).
Udugama, B. et al. Kuzindikira kwa COVID-19: Matenda ndi Zida Zoyesera. ACS Nano 14(4), 3822–3835 (2020).
Syed S. et al. Kukhazikitsidwa kwa College of Pathologists of Eastern, Central and Southern Africa - Regional School of Pathology of the Middle East and South Africa. Africa. J. Labu. mankhwala. 9(1), 1-8 (2020).
Ethiopia Institute of Public Health, Federal Ministry of Health. Interim National Strategy and Guidance for Laboratory Diagnosis of COVID-19. https://ephi.gov.et/images/novel_coronavirus/EPHI_PHEOC_COVID-19_Laboratory_Diagnosis_Eng.pdf (yofikira pa Ogasiti 12, 2020) (EPHI, 2020).
Woloshin, S., Patel, N. & Kesselheim, AS Mayeso abodza azovuta za matenda a SARS-CoV-2 ndi zotsatira zake. Woloshin, S., Patel, N. & Kesselheim, AS Mayeso abodza azovuta za matenda a SARS-CoV-2 ndi zotsatira zake.Voloshin S., Patel N. ndi Kesselheim AS Mayeso abodza a matenda a SARS-CoV-2 ndi zotsatira zake.Voloshin S., Patel N. ndi Kesselheim AS Mayeso abodza oyambitsa matenda a SARS-CoV-2. N. Enga. J. Mankhwala. 383(6), e38 (2020).
Mouliou, DS & Gourgoulianis, KI Nkhani zabodza komanso zabodza za COVID-19: Njira zopewera kupuma ndi kasamalidwe, katemera, ndi malingaliro ena. Mouliou, DS & Gourgoulianis, KI Nkhani zabodza komanso zabodza za COVID-19: Njira zopewera kupuma ndi kasamalidwe, katemera, ndi malingaliro ena. Mouliou, DS & Gourgoulianis, KI Ложноположительные ndi ложноотрицательные kulimbana ndi COVID-19: респираторная профилактика и сратегии лечения, ваиякишина перспективы. Mouliou, DS & Gourgoulianis, KI Nkhani zabodza komanso zabodza za COVID-19: njira zopewera kupuma ndi chithandizo, katemera ndi njira yakutsogolo.Muliu, DS ndi Gurgulianis, KI Nkhani zabodza komanso zabodza za COVID-19: njira zopewera kupuma ndi chithandizo, katemera ndi njira yopita patsogolo. Katswiri Reverend Respire. mankhwala. 15(8), 993–1002 (2021).
Mouliou, DS, Ioannis, P. & Konstantinos, G. COVID-19 matenda mu dipatimenti yadzidzidzi: Kuwona mtengo koma kutaya nkhalango. Mouliou, DS, Ioannis, P. & Konstantinos, G. COVID-19 matenda mu dipatimenti yadzidzidzi: Kuwona mtengo koma kutaya nkhalango.Mouliou, DS, Ioannis, P. ndi Konstantinos, G. COVID-19 Kuzindikira mu Dipatimenti Yowopsa: Onani Mtengo, Taya Nkhalango.Muliou DS, Ioannis P., ndi Konstantinos G. Matenda a COVID-19 M'zipinda Zadzidzidzi: Palibe Nkhalango Yokwanira ya Mitengo. Kuwoneka. mankhwala. J. https://doi.org/10.1136/emermed-2021-212219 (2022).
Degli-Angeli, E. et al. Kutsimikizika ndi Kutsimikizika kwa Ntchito Yowunika ndi Yachipatala ya Abbott RealTime SARS-CoV-2 Assay. J. Zachipatala. Kachilombo. 129, 104474. https://doi.org/10.1016/j.jcv.2020.104474 (2020).
Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Fananizani ma seti oyambira asanu ochokera kumadera osiyanasiyana a ma genome a COVID-19 kuti azindikire kachilombo ka HIV ndi RT-PCR wamba. Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Kuyerekeza kwa magawo asanu oyambirira ochokera kumadera osiyanasiyana a genome a COVID-19 kuti azindikire kachilombo ka HIV ndi RT-PCR wamba.Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. ndi Aflatunyan, B. Kuyerekeza kwa magulu asanu oyambira ochokera kumadera osiyanasiyana a COVID-19 genome pozindikira matenda a virus ndi RT-PCR wamba. 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迩组-PC检测病毒感染. Mollaei, HR, Afshar, AA, Kalantar-Neyestanaki, D., Fazlalipour, M. & Aflatoonian, B. Kuyerekeza madera 5 osiyanasiyana amtundu wa COVID-19 pozindikira matenda a virus ndi RT-PCR wamba.Mollaei HR, Afshar AA, Kalantar-Neyestanaki D, Fazlalipour M. ndi Aflatunyan B. Kuyerekeza kwa magulu asanu oyambira ochokera kumadera osiyanasiyana a COVID-19 genome pozindikira matenda a virus ndi RT-PCR wamba.Iran. J. Microbiology. 12 (3), 185 (2020).
Goertzer, I. et al. Zotsatira zoyambilira za pulogalamu yowunika zamtundu wakunja zamtundu wa SARS-CoV-2 genome sequence. J. Zachipatala. Kachilombo. 129, 104537. https://doi.org/10.1016/j.jcv.2020.104537 (2020).
Wang, M. et al. Kuwunika Kuwunika kwa Mphamvu kwa Zida Zisanu za RT-PCR za Severe Acute Respiratory Syndrome Coronavirus 2. J. Clinical. labotale. anus. 35(1), e23643 (2021).
Wang B. et al. Kuunikira kwa zida zisanu ndi ziwiri zomwe zikupezeka pamalonda za SARS-CoV-2 RNA ku China kutengera zenizeni zenizeni za polymerase chain reaction (PCR). zachipatala. Chemical. labotale. mankhwala. 58(9), e149–e153 (2020).
van Casteren, PB et al. Kuyerekeza kwa zida zisanu ndi ziwiri zamalonda za RT-PCR COVID-19. J. Zachipatala. Kachilombo. 128, 104412 (2020).
Lu, Yu, et al. Kuyerekeza kwa magwiridwe antchito a zida ziwiri za PCR pozindikira ma SARS-CoV-2 nucleic acid. J. Zachipatala. labotale. anus. 34(10), e23554 (2020).
Lefart, PR, etc. Kafukufuku woyerekeza anayi SARS-CoV-2 nucleic acid amplification test (NAAT) nsanja anasonyeza kuti ID TSOPANO ntchito inali yonyozeka kwambiri malinga ndi wodwala ndi chitsanzo mtundu. matenda. microbiology. Kupatsirana. diss. 99(1), 115200 (2021).
Molekyu ya Abbott. Abbott real-time SARS-CoV-2 phukusi lowunikira. https://www.molecular.abbott/us/en/products/infectious-disease/RealTime-SARS-CoV-2-Assay. 1-12. (Kuyambira pa Ogasiti 10, 2020) (2020).
Klein, S. et al. Kudzipatula kwa SARS-CoV-2 RNA pogwiritsa ntchito mikanda ya maginito kuti izindikire mwachangu kwambiri ndi RT-qPCR ndi RT-LAMP. Virus 12(8), 863 (2020).