Ayyukan gwaje-gwajen haɓaka acid nucleic guda huɗu don gano SARS-CoV-2 a Habasha

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Tun bayan barkewar cutar coronavirus ta 2019 (COVID-19), yawancin gwajin haɓaka haɓakar acid nucleic acid (NAATs) an haɓaka su a duk duniya kuma sun zama daidaitattun ƙididdiga. Ko da yake an haɓaka gwaje-gwaje da yawa cikin sauri kuma an yi amfani da su ga gwaje-gwajen bincike na dakin gwaje-gwaje, ba a tantance aikin waɗannan gwaje-gwajen ba a cikin saituna daban-daban. Don haka, wannan binciken ya yi niyya don kimanta aikin Abbott SARS-CoV-2, Daan Gene, BGI, da Sansure Biotech assays ta amfani da Ma'aunin Magana (CRS). An gudanar da binciken ne a Cibiyar Kiwon Lafiyar Jama'a ta Habasha (EPHI) daga 1 zuwa 30 ga Disamba 2020. An fitar da samfuran nasopharyngeal guda 164 ta amfani da ƙaramin kit ɗin QIAamp RNA da tsarin tsarin samfurin Abbott DNA. Daga cikin samfurori 164, 59.1% sun kasance masu inganci kuma 40.9% ba su da kyau ga CRS. Matsayin Sansure Biotech ya yi ƙasa sosai idan aka kwatanta da CRS (p <0.05). Matsayin Sansure Biotech ya yi ƙasa sosai idan aka kwatanta da CRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p <0,05). Ingantattun sakamakon Sansure Biotech sun yi ƙasa sosai idan aka kwatanta da 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 yana da ƙarancin sakamako masu inganci idan aka kwatanta da CRS (p <0.05).Yarjejeniyar gabaɗaya na ƙididdigar huɗun ita ce 96.3-100% idan aka kwatanta da CRS. Bugu da ƙari ga ƙarancin ƙima na Sansure Biotech assay, aikin gwaje-gwajen guda huɗu ya kusan kwatanta. Don haka, ƙididdigar Sansure Biotech [Bincike Kawai (RUO)] yana buƙatar ƙarin ingantaccen amfani da shi a Habasha. A ƙarshe, ya kamata a yi la'akari da ƙarin bincike don kimanta ƙima tare da da'awar masana'anta da suka dace.
Gwajin dakin gwaje-gwaje wani bangare ne na Tsarin Dabarun Hukumar Lafiya ta Duniya (WHO) don Cutar Coronavirus 2019 (COVID-19) Shirye-shirye da Amsa (SPRP). WHO ta ba da shawarar cewa kasashe suna buƙatar haɓaka ƙarfin dakin gwaje-gwaje don haɓaka shirye-shirye, gudanar da shari'o'in da ya dace, yin taka-tsantsan da saurin mayar da martani ga ƙalubalen lafiyar jama'a. Wannan yana nuna cewa aikin dakin gwaje-gwaje shine mabuɗin don siffanta cututtuka da cututtukan cututtukan cututtukan da ke tasowa da kuma shawo kan yaduwar su.
Binciken COVID-19 yana buƙatar bayanan annoba da na likitanci, alamun mutum/alamu, da bayanan rediyo da dakin gwaje-gwaje2. Tun lokacin da aka ba da rahoton bullar cutar ta COVID-19 a birnin Wuhan na kasar Sin, an samar da gwaje-gwajen kara karfin acid na kasuwanci da yawa (NAATs) a duk duniya. An yi amfani da aikin jujjuyawar juzu'i na ainihin-lokaci polymerase sarkar dauki (rRT-PCR) azaman tsarin yau da kullun kuma daidaitaccen hanya don ganewar dakin gwaje-gwaje na kamuwa da cuta mai tsanani na numfashi 2 (SARS-CoV-2) 3. Gano kwayoyin halitta na SARS-CoV-2 yawanci ya dogara ne akan N (jinin furotin nucleocapsid), E (jini na furotin ambulan), da RdRp (genes RNA polymerase gene) mai dogaro da RdRp a cikin ORF1a/b (budadden firam ɗin karatun 1a/b) . gene) yankin da aka gano daga kwayar cutar kwayar cuta. Ana la'akari da su a matsayin manyan yankuna da aka adana a cikin kwayar cutar kwayar cutar kwayar cutar kwayar cuta4. Daga cikin wadannan kwayoyin halittar, kwayoyin halittar RdRp da E suna da karfin ganowa na nazari, yayin da kwayar halittar N tana da karancin hankali na nazari5.
Ayyukan gwaje-gwaje na PCR na iya bambanta dangane da dalilai daban-daban kamar: reagents hakar, haɓakawa / gano reagents, hanyar hakar, ingancin injin PCR da sauran kayan aikin. Ya zuwa Afrilu 2020, fiye da na'urorin bincike daban-daban 48 daga ƙasashe tara sun karɓi Izinin Amfani da Gaggawa (EUA) don gwajin COVID-196. A Habasha, ana amfani da dandamali sama da 14 na ainihin PCR don gano PCR na SARS-CoV-2 a cibiyoyin kiwon lafiyar jama'a 26, gami da ABI 7500, Abbott m2000, Roche 48000 da Quant-studio7. Bugu da kari, ana samun na'urorin gwajin PCR daban-daban, kamar gwajin Daan Gene, gwajin Abbott SARS-CoV-2, gwajin Sansure Biotech, da gwajin SARS-CoV-2 BGI. Kodayake rRT-PCR tana da hankali sosai, wasu marasa lafiya tare da COVID-19 suna ba da rahoton sakamako mara kyau na ƙarya saboda ƙarancin kwafin ribonucleic acid (RNA) a cikin samfura saboda tarin da bai dace ba, jigilar kaya, ajiya da sarrafawa, da gwajin dakin gwaje-gwaje. yanayi da ayyukan ma'aikata8. Bugu da kari, samfurin ko sarrafa kuskure, saitin kewayawa (Ct), da haɗin kai tare da sauran ƙwayoyin cuta na ƙwayoyin cuta ko SARS-CoV-2 RNA marasa aiki / saura na iya haifar da sakamako mai kyau na ƙarya a cikin ƙididdigar rRT-PCR9. Don haka, a bayyane yake cewa gwaje-gwajen PCR na iya gano masu ɗauke da gutsuttsuran kwayoyin halitta, saboda ba za su iya ma bambance tsakanin ƙwayoyin ƙwayoyin cuta masu aiki da gaske ba, don haka gwaje-gwajen na iya gano masu ɗaukar hoto ne kawai ba marasa lafiya ba10. Sabili da haka, yana da mahimmanci don tantance aikin bincike ta amfani da daidaitattun hanyoyin a cikin saitin mu. Duk da cewa ana samun magungunan NAAT da yawa a Cibiyar Kiwon Lafiyar Jama'a ta Habasha (EPHI) da kuma ko'ina cikin ƙasar, har yanzu ba a sami rahoton kwatancen ingancinsu ba. Don haka, wannan binciken ya yi niyya don kimanta aikin kwatankwacin kayan aikin kasuwanci don gano SARS-CoV-2 ta rRT-PCR ta amfani da samfuran asibiti.
Adadin mahalarta 164 da ake zargin COVID-19 an haɗa su cikin wannan binciken. Yawancin samfurori sun fito ne daga cibiyoyin kulawa (118/164 = 72%), yayin da sauran 46 (28%) mahalarta sun kasance daga cibiyoyin marasa magani. Daga cikin mahalarta da ba a kula da su ba a cibiyar, 15 (9.1%) suna da shari'o'in da ake zargin asibiti da kuma 31 (18.9%) suna da lambobin da aka tabbatar. Mahalarta casa'in da uku (56.7%) maza ne, kuma ma'anar (± SD) shekarun mahalarta shine 31.10 (± 11.82) shekaru.
A cikin wannan binciken, an ƙaddara ƙimar inganci da mara kyau na gwaje-gwaje huɗu na COVID-19. Don haka, ingantattun ƙimar Abbott SARS-CoV-2 assay, Daan Gene 2019-nCoV assay, SARS-CoV-2 BGI assay, da Sansure Biotech 2019-nCoV assay sun kasance 59.1%, 58.5%, 57.9% da 55.5% bi da bi. . Makiyoyin ma'auni masu inganci da mara kyau (CRS) sune 97 (59.1%) da 67 (40.9%), bi da bi (Table 1). A cikin wannan binciken, ma'anar CRS ta dogara ne akan ka'idar "kowace mai kyau", wanda daga cikin sakamakon gwaji guda hudu, sakamakon gwaji biyu ko fiye da suka ba da sakamako iri ɗaya an dauke su gaskiya ne ko mara kyau.
A cikin wannan binciken, mun sami yarjejeniya mara kyau (NPA) na 100% (95% CI 94.6-100) don duk nazarin idan aka kwatanta da CRS. Binciken Sansure Biotechnology ya nuna ƙaramin PPA na 93.8% (95% CI 87.2-97.1) da kuma binciken Daan Gene 2019-nCoV yana da cikakkiyar yarjejeniya ta 99.4% (95% CI 96.6-99.9). Sabanin haka, gaba ɗaya yarjejeniya tsakanin SARS-CoV-2 BGI assay da Sansure Biotech 2019-nCoV assay ya kasance 98.8% da 96.3%, bi da bi (Table 2).
Cohen's kappa coefficient na yarjejeniya tsakanin CRS da Abbott SARS-CoV-2 sakamakon assay ya kasance cikakke (K = 1.00). Hakanan, ƙimar kappa na Cohen da Daan Gene 2019-nCoV, SARS-CoV-2 BGI, da Sansure Biotech 2019-nCoV suka gano sun yi daidai da CRS (K ≥ 0.925). A cikin wannan kwatancen kwatancen, gwajin chi-square (gwajin McNemar) ya nuna cewa sakamakon binciken Sansure Biotech 2019-nCoV ya bambanta sosai da sakamakon CRS (p = 0.031) (Table 2).
Kamar yadda aka nuna a cikin Fig.1 kashi mafi ƙarancin ƙimar Ct (<20 Ct) na Abbott SARS-CoV-2 assay (haɗe RdRp da N gene) ya kasance 87.6% da ORF1a/b gene Ct darajar Sansure Biotech 2019-nCoV assay ya nuna cewa adadin ƙarancin Ƙimar Ct (<20 Ct) ya kasance 50.3% kuma babban darajar Ct (36-40 Ct) ya canza zuwa +3.2%. 1 kashi mafi ƙarancin ƙimar Ct (<20 Ct) na Abbott SARS-CoV-2 assay (haɗe RdRp da N gene) ya kasance 87.6% da ORF1a/b gene Ct darajar Sansure Biotech 2019-nCoV assay ya nuna cewa adadin ƙarancin Ƙimar Ct (<20 Ct) ya kasance 50.3% kuma babban darajar Ct (36-40 Ct) ya canza zuwa +3.2%.Kamar yadda aka nuna a cikin Fig.1, процент наименьшего значения Ct (< 20 Ct) анализа Abbott SARS-CoV-2 (комбинированный ген RdRp и N) составил 87, 6% ORF1a/b анализа Sansure Biotech 2019-nCoV показало что процент низкого значения Ct (< 20 Ct) составлял 50,3%, а высначкое 4 kashi 3.2%. 1, adadin mafi ƙarancin ƙimar Ct (< 20 Ct) na Abbott SARS-CoV-2 (haɗe-haɗe gene RdRp da N) ya kasance 87.6%, kuma ƙimar Ct na ORF1a/b nazarin halittar Sansure Biotech 2019-nCoV ya nuna. cewa yawan ƙananan ƙimar Ct (<20 Ct) ya ƙidaya 50.3%, kuma babban darajar Ct (36-40 Ct) ya kai 3.2%.Abbott SARS-CoV-2 检测（结合RdRp 和N 基因）的最低Ct 值百分比（< 20 Ct） 8Ssure ​​Biotechn检测的ORF1a/b 基因Ct 值显示低Ct 值(< 20 Ct) 的百分比为50.3%，高Ct 值(36-40 Ct) Kamar yadda aka nuna a cikin Hoto 1, mafi ƙarancin ƙimar Ct (<20 Ct) na gwajin Abbott SARS-CoV-2 (haɗin RdRp da N gene) shine 87.6%, ƙimar ORF1a/b gene Ct na gwajin Sansure Biotech 2019-nCoV yana nuna ƙananan Ct值 (< 20 Ct) 的 kashi 50.3% ne, 高Ct值 (36-40 Ct) 的 kashi 3.2%. Как показано на рисунке 1, анализ Abbott SARS-CoV-2 (сочетающий гены RdRp и N) размере 87,6%, а значение Ct гена ORF1a/b исследовании Sansure Biotech 2019- Анализ nCoV показал низкий Ct. Kamar yadda aka nuna a cikin Hoto 1, Abbott SARS-CoV-2 assay (haɗe da RdRp da N genes) yana da mafi ƙarancin ƙimar Ct (<20 Ct) a 87.6%, yayin da ƙimar Ct na ORF1a/b gene a cikin Sansure Nazarin Biotech 2019 - Binciken nCoV ya nuna ƙarancin Ct. Процент значений (<20 Ct) составил 50.3%, а процент высоких значений Ct (36-40 Ct) составил 3,2%. Adadin ƙimar (<20 Ct) shine 50.3%, kuma adadin manyan ƙimar Ct (36-40 Ct) shine 3.2%.Gwajin Abbott SARS-CoV-2 B ya yi rikodin ƙimar Ct sama da 30. A gefe guda, akan BGI SARS-CoV-2 assay ORF1a/b gene yana da ƙimar Ct mai girma (> 36 Ct) kashi 4% (Fig. 1). A gefe guda, akan BGI SARS-CoV-2 assay ORF1a/b gene yana da ƙimar Ct mai girma (> 36 Ct) kashi 4% (Fig. 1). Сдругой стороны, в анализе BGI SARS-CoV-2 ген ORF1a/b имел высокое значение Ct (> 36 Ct), процент которого сир. A gefe guda, a cikin nazarin BGI SARS-CoV-2 gene ORF1a/b yana da ƙimar Ct mai girma (> 36 Ct), adadin wanda ya kasance 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） A gefe guda, a cikin gano BGI SARS-CoV-2, adadin ORF1a/b gene tare da ƙimar Ct mai girma (> 36 Ct) shine 4% (Hoto 1). Сдругой стороны, в анализе BGI SARS-CoV-2 процент генов ORF1a/b с высокими значениями Ct (> 36 Ct) составил 4% (ри). A gefe guda, a cikin binciken BGI SARS-CoV-2, yawan adadin kwayoyin ORF1a/b tare da ƙimar Ct masu girma (> 36 Ct) shine 4% (Fig. 1).
A cikin wannan binciken, mun ɗauki samfuran nasopharyngeal 164. Don kowane nau'in tantancewa, warewar RNA da haɓakawa an yi su ta amfani da hanyoyin da kayan aikin da masana'antun suka ba da shawarar.
Wannan binciken ya nuna cewa gwajin Abbott na SARS-CoV-2 yana da aikin ganowa iri ɗaya da CRS, tare da tabbataccen 100%, mara kyau, da haɗin gwiwa gabaɗaya. Yarjejeniyar kappa ta Cohen shine 1.00, yana nuna cikakkiyar yarjejeniya tare da CRS. Wani irin binciken da Jami'ar Washington ta yi a Amurka ya gano cewa gabaɗayan hankali da ƙayyadaddun gwajin Abbott na SARS-CoV-2 ya kasance 93% da 100%, bi da bi, idan aka kwatanta da ƙayyadaddun gwaje-gwaje (LDA) na CDC. . 11. The Abbott SARS-CoV-2 tsarin ganowa ya dogara ne a kan lokaci guda gano kwayoyin N da RdRp, kamar yadda duka kwayoyin halitta sun fi damuwa, rage girman rashin kuskure12. Wani bincike a Vienna, Ostiriya ya kuma nuna cewa manyan ɗimbin samfuran hakar da gano abubuwan ganowa sun rage tasirin dilution da haɓaka haɓakar ganowa13. Don haka, cikakkiyar madaidaicin Abbott don gwajin SARS-CoV-2 na iya haɗawa da tsarin gano dandamali wanda ke gano ƙwayoyin halittar haɗin gwiwa lokaci guda, yana fitar da samfuran samfuri masu yawa (0.5 ml), kuma yana amfani da adadi mai yawa na eluent (40 µl).
Sakamakonmu ya kuma nuna cewa aikin gano gwajin kwayoyin halittar Daan ya kusan zama daidai da na CRS. Wannan ya yi daidai da binciken14 da aka gudanar a Jami'ar Anhui a Huainan, China, da da'awar masana'anta na 100% tabbataccen yarjejeniya. Duk da rahotannin tabbataccen sakamako, samfurin guda ɗaya ya kasance mara kyau bayan sake gwadawa iri ɗaya, amma yana da inganci a cikin Abbott SARS-CoV-2 da Sansure Biotech nCoV-2019 assays. Wannan yana nuna cewa za'a iya samun sauye-sauye a cikin sakamako a cikin nau'o'in ƙididdiga daban-daban. Duk da haka, a cikin binciken da aka gudanar a China15, sakamakon gwajin Daan Gene ya bambanta sosai (p <0.05) idan aka kwatanta da ƙayyadaddun bayanan binciken su. Duk da haka, a cikin binciken da aka gudanar a China15, sakamakon gwajin Daan Gene ya bambanta sosai (p <0.05) idan aka kwatanta da ƙayyadaddun bayanan binciken su. Тем не менее, в исследовании, проведенном в Китае15, результат анализа Daan Gene значительно отличался, 5 <p laboratornogo эtalonnogo analiza. Koyaya, a cikin binciken da aka yi a China15, sakamakon binciken Daan Gene ya bambanta sosai (p <0.05) daga binciken binciken binciken su.然而, 国进行国进行 的 研究 中 15, 大安基因检测 的 的 的 参考检测相比参考检测相比 (p <0.05)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的参室定义的参考检测縭参考检测績来0. Однако в исследовании, проведенном в Китае15, результаты генетического теста Daan значительно оть (5) <0) sravnenyyu с егo эtalonnыm labortornыm testom. Koyaya, a cikin binciken da aka yi a China15, sakamakon gwajin kwayoyin halittar Daan ya bambanta sosai (p <0.05) idan aka kwatanta da gwajin dakin gwaje-gwaje.Wannan bambance-bambancen na iya kasancewa saboda azancin gwajin tunani don gano SARS-CoV-2, kuma ƙarin karatun na iya zama mahimmanci don tantance dalilin.
Bugu da kari, bincikenmu ya kimanta aikin kwatankwacin aikin SARS-CoV-2 BGI tare da CRS, yana nuna kyakkyawar yarjejeniya mai inganci (PPA = 97.9%), yarjejeniya mara kyau (NPA = 100%), da yarjejeniyar kashi gaba daya ta jinsi ( OPA). ). = 98.8%). Ƙimar Cohen's Kappa sun nuna kyakkyawar yarjejeniya (K = 0.975). Nazarin da aka yi a Netherlands16 da China15 sun nuna sakamako daidai gwargwado. Gwajin SARS-CoV-2 BGI gwajin ganowa ce guda ɗaya (ORF1a/b) ta amfani da haɓakawa / ganowa 10 µl. Duk da kyakkyawar yarjejeniya ta ƙididdiga tare da sakamakon binciken mu, binciken ya rasa samfurori biyu masu kyau (1.22%) na jimlar samfurin. Wannan na iya samun babban tasiri na asibiti don haɓakar watsawa a duka matakan haƙuri da na al'umma.
Wani kwatancen bincike da aka haɗa a cikin wannan binciken shine Sansure Biotech nCoV-2019 rRT-PCR (RUO); jimlar yawan wasan ya kasance 96.3%. Ƙarfin yarjejeniya kuma an ƙayyade ƙimar Cohen's Kappa, wanda shine 0.925, yana nuna cikakkiyar yarjejeniya tare da CRS. Bugu da kari, sakamakonmu ya yi daidai da nazarce-nazarcen da aka gudanar a Jami'ar Kudu ta Tsakiya da ke birnin Changsha na kasar Sin, da kuma a Sashen dakunan gwaje-gwaje na asibitin jama'ar Liuzhou, birnin Liuzhou na kasar Sin17. Ko da yake an yi rikodin ƙididdiga mai kyau na sama, gwajin chi-square (gwajin MacNemar) ya nuna cewa sakamakon Sansure Biotech assay yana da bambanci mai mahimmanci idan aka kwatanta da CRS (p <0.005). Ko da yake an yi rikodin ƙididdiga mai kyau na sama, gwajin chi-square (gwajin MacNemar) ya nuna cewa sakamakon Sansure Biotech assay yana da bambanci mai mahimmanci idan aka kwatanta da CRS (p <0.005). Несмотря на то, что было зафиксировано указанное выше. (критерий Макнемара) показал, что результат анализа Sansure Biotech Ko da yake an yi rikodin kyakkyawar yarjejeniyar ƙididdiga ta sama, gwajin chi-square (gwajin McNemar) ya nuna cewa sakamakon Sansure Biotech assay yana da bambanci mai mahimmanci idan aka kwatanta da 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相比具有统计学显着差异（p <0.005）。尽管 记录 了 上述 良好 统计 一致性 ， 但 检验 （（macnemar相比 具有 显着 （（p <0.005））） Несмотря на отмеченое выше хорошее статистическое соответствие статистически значимую разницу (p <0,005) между анализом Sansure Biotech и CRS. Duk da kyakkyawar yarjejeniyar ƙididdiga da aka ambata a sama, gwajin chi-square (gwajin McNemar) ya nuna bambanci mai mahimmanci (p <0.005) tsakanin Sansure Biotech assay da CRS.An samo samfurori shida (3.66%) a matsayin rashin kuskure idan aka kwatanta da CRS (Ƙarin Teburin 1); wannan yana da matukar muhimmanci, musamman idan aka yi la’akari da yadda kwayar cutar ke yaduwa. Bayanan da ke sama kuma suna goyan bayan wannan ƙananan ƙimar ganowa15.
A cikin wannan binciken, an ƙaddara ƙimar Ct don kowane ƙima da dandamali daban-daban, tare da mafi ƙarancin ƙimar Ct da aka ruwaito a cikin Abbott SARS-CoV-2 assay. Wannan sakamakon na iya kasancewa yana da alaƙa da tsarin gwajin kwayoyin halittar Abbott na lokaci guda don gano SARS-CoV-2. Sabili da haka, bisa ga Hoto 1, 87.6% na sakamakon Abbott SARS-CoV-2 yana da ƙimar Ct a ƙasa da 20. Ƙananan adadin sakamakon samfurin (12.4%) ne kawai ke cikin kewayon 20-30. Ba a rubuta ƙimar Ct sama da 30 ba. Baya ga amfani da Abbott na tsarin gwajin kwayoyin halittar SARS-CoV-2 panel, wannan sakamakon na iya kasancewa yana da alaƙa da ƙarancin ganowa (kwafin 32.5 RNA / ml) 18, wanda ya ninka sau uku ƙasa da ƙarancin kamfanin na kwafin 100 RNA. /ml. ml) 19.
Wannan binciken yana da wasu iyakoki: na farko, ba mu da daidaitattun hanyoyin / hanyoyin magana [kamar kwayar cutar hoto ko wasu gwaje-gwajen dakin gwaje-gwaje (LDA)] saboda rashin albarkatu. Na biyu, duk samfuran da aka yi amfani da su a cikin wannan binciken sune swabs na nasopharyngeal, yayin da sakamakon bai dace da wasu nau'in samfurori ba, kuma na uku, girman samfurin mu ya kasance ƙananan.
Wannan binciken ya kwatanta aikin gwaje-gwajen rRT-PCR guda huɗu don SARS-CoV-2 ta amfani da samfuran nasopharyngeal. Duk gwaje-gwajen ganowa sun yi kusan kwatankwacin aiki, ban da Sansure Biotech assay. Bayan haka, an gano ƙarancin ƙima a cikin binciken Sansure Biotech idan aka kwatanta da CRS (p <0.05). Bayan haka, an gano ƙarancin ƙima a cikin binciken Sansure Biotech idan aka kwatanta da CRS (p <0.05). Кроме того, в тесте Sansure Biotech был выявлен низкий процент положительных результатов по сравнению с CRS (p <0,0). Bugu da ƙari, gwajin Sansure Biotech ya nuna ƙarancin sakamako mai kyau idan aka kwatanta da 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 имел более низкий уровень положительных результатов по сравнению с CRS (p <0,05). Bugu da kari, Sansure Biotech assay yana da ƙarancin inganci idan aka kwatanta da CRS (p <0.05).Binciken Sansure Biotech nCoV-2019 (RUO) na PPA, NPA da yarjejeniya gabaɗaya ya wuce 93.5% tare da ƙarfin Cohen Kappa na ƙimar yarjejeniya na 0.925. A ƙarshe, Sansure Biotech Assay (RUO) yana buƙatar ƙarin tabbaci don amfani a Habasha, kuma ya kamata a yi la'akari da ƙarin bincike don kimanta da'awar daga masana'anta guda ɗaya.
An gudanar da tsarin nazarin kwatancen a cibiyoyin lafiya guda hudu a Addis Ababa, Asibitin Eka Kotebe, Cibiyar Kula da Majami'a ta Millennium, Asibitin Tunawa da Zewooditu, da Asibitin Kwararru na Tuberculosis na St. Peter. An tattara bayanan tsakanin 1 ga Disamba zuwa 31 ga Disamba, 2020. An zaɓi wuraren kiwon lafiya don wannan binciken da gangan bisa la'akari da yawan adadinsu da kuma kasancewar manyan cibiyoyin jiyya a cikin birni. Hakanan, kayan aikin, gami da ABI 7500 da Abbott m2000 na ainihin kayan aikin PCR, an zaɓi su bisa ga shawarwarin masana'antun NAAT reagent, kuma an zaɓi na'urorin gano PCR guda huɗu don wannan binciken, kamar yadda yawancin dakunan gwaje-gwaje a Habasha ke amfani da aƙalla aƙalla. hudu daga cikinsu. Gwajin Gene, gwajin Abbott SARS-CoV-2, gwajin Sansure Biotech, da gwajin SARS-CoV-2 BGI da aka yi yayin binciken).
An yi gwajin SARS-CoV-2 daga 1 zuwa 30 ga Disamba 2020 ta amfani da 3 ml na Matsakaicin Sufuri na Viral (VTM) (Fasaha ta Mu'ujiza, Shenzhen, China) daga mutanen da ke kan bincike don COVID-19 da ake magana da su EPHI. An tattara samfuran nasopharyngeal daga ƙwararrun masu tattara samfuran kuma an aika zuwa EPHI cikin fakiti uku. Kafin warewa acid nucleic, kowane samfurin ana sanya shi lambar shaida ta musamman. Ana yin hakar daga kowane samfurin nan da nan da isowa ta hanyar amfani da jagora da hanyoyin hakar atomatik. Don haka, don hakar atomatik na Abbott m2000, 1.3 ml (ciki har da 0.8 ml matattu ƙarar da 0.5 ml hakar mashiga ƙarar) na samfurin an ciro daga kowane samfurin kuma an wuce ta hanyar Abbott DNA Samfurin Shirye-shiryen Tsarin (Abbott Molecular Inc. des Plaines). IL, Amurka). ) Batch na 96 [samfurori 92, sarrafawar ganowa biyu da na'urori marasa samfuri guda biyu (NTC)] an haɗa su a cikin tsarin gabaɗaya (dawowa da ganowa) na zagaye biyu na SARS-CoV-2 (EUA) a cikin ainihin lokaci. hakar ma'adinai. Hakazalika, don hakar hannu, yi amfani da samfurori iri ɗaya (don hakar atomatik da ganowa). Don haka, a duk tsawon aikin, an fitar da samfuran 140 µl kuma an fitar da su ta amfani da QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Jamus) a cikin batches na 24 (ciki har da samfuran 20, sarrafawar assay biyu da NTCs biyu) sama da zagaye tara. Abubuwan da aka fitar da hannu an haɓaka su kuma an gano su ta amfani da ABI 7500 thermal cycler ta amfani da SARS-CoV-2 BGI assay, Daan Gene assay, da Sansure Biotech assay.
Warewa ta atomatik da tsarkakewa na SARS-CoV-2 viral RNA yana bin ka'idodin bead na maganadisu ta amfani da samfuran samfuran Abbott DNA. Rashin kunna samfurori da solubilization na ƙwayoyin cuta ana aiwatar da su ta hanyar amfani da wanki mai ƙunshe da guanidine isothiocyanate don cire furotin kuma ya hana RNase aiki. Ana raba RNA daga furotin ta hanyar rabuwa mai ƙarfi ta amfani da silica, watau gishirin guanidinium da alkaline pH na ma'aunin lysis suna haɓaka ɗaurin acid nucleic zuwa silica (SiO2). Matakin kurkura yana kawar da sauran sunadaran da tarkace don samar da ingantaccen bayani. RNA mai bayyanawa an keɓe shi daga silica tushen microparticles ta amfani da filin maganadisu na kayan aiki20,21. A gefe guda, keɓewar hannu da tsarkakewa na RNA ana aiwatar da su ta hanyar ginshiƙin juzu'i ta hanyar amfani da centrifugation maimakon madaidaicin maganadisu da rabuwar ƙananan ƙwayoyin cuta daga eluent.
Gwajin Ganowar Abbott Real-Time SARS-CoV-2 (Abbott Molecular, Inc.) an yi shi bisa ga umarnin masana'anta, wanda ya karɓi EUA19,22 daga WHO da FDA. A cikin wannan ka'ida, samfurin rashin kunnawa kafin cirewa an yi shi a cikin wanka na ruwa a 56 ° C na 30 min. Bayan rashin kunna ƙwayoyin cuta, an yi hakar acid nucleic akan kayan aikin Abbott m2000 SP daga 0.5 ml VTM ta amfani da tsarin shirye-shiryen samfurin DNA na Abbott m2000. bisa ga masana'anta. An yi haɓakawa da ganowa ta amfani da kayan aikin Abbott m2000 RT-PCR, kuma an yi gano dual don ƙwayoyin RdRp da N. ROX) da VIC P (dini na mallakar mallaka) don yin niyya da gano abubuwan sarrafawa na ciki, ba da izinin gano lokaci guda na samfuran haɓakawa 19.
Hanyar gano haɓakawa na wannan kit ɗin ya dogara ne akan fasahar RT-PCR mataki ɗaya. An zaɓi kwayoyin ORF1a/b da N a matsayin yankunan da aka kiyaye su ta Daan Gene Technology don gano haɓaka yankin da aka yi niyya. Takamaiman firamare da bincike mai kyalli (binciken kwayoyin halittar N da aka yiwa lakabi da FAM, ORF1a/b binciken da aka yiwa lakabi da VIC) an tsara su don gano SARS-CoV-2 RNA a cikin samfuran. An shirya eluent na ƙarshe da haɗe-haɗe ta hanyar ƙara 5 µl na eluent zuwa 20 µl na mahaɗin maigida zuwa ƙarar ƙarshe na 25 µl. An yi haɓakawa da ganowa lokaci guda akan kayan aikin ABI 750024 na ainihin lokaci na PCR.
An gano kwayoyin ORF1a/b da N ta amfani da Sansure Biotech nCoV-2019 Nucleic Acid Diagnostic Kit (gano PCR mai kyalli). Shirya ƙayyadaddun bincike don kowane nau'in manufa ta hanyar zaɓar tashar FAM don yankin ORF1a/b da tashar ROX don kwayar halittar N. Zuwa wannan kit ɗin kit ɗin, ana ƙara eluent da master mix reagents kamar haka: shirya 30 µl master mix reagent da 20 µl eluted samfurin don ganowa / haɓakawa. An yi amfani da PCR ABI 750025 na ainihi don haɓakawa/ganewa.
Gwajin SARS-CoV-2 BGI shine kayan kyalli na gaske na rRT-PCR don gano cutar COVID-19. Yankin da aka yi niyya yana cikin yankin ORF1a/b na kwayoyin halittar SARS-CoV-2, wanda shine hanyar gano kwayoyin halitta guda daya. Bugu da kari, kwayar halittar dan adam β-actin wata kwayar halitta ce da aka tsara ta cikin gida. Ana shirya mahaɗin maigidan ta hanyar haɗa 20 µl na reagent master mix da 10 µl na samfurin RNA da aka fitar a cikin rijiyar faranti26. An yi amfani da kayan aikin PCR na gaske na ABI 7500 don haɓakawa da ganowa. Duk haɓakar acid nucleic, yanayin tafiyar da PCR don kowane ƙima, da fassarar sakamako an yi su bisa ga umarnin masana'anta (Table 3).
A cikin wannan kwatancen kwatancen, ba mu yi amfani da ma'aunin ma'auni ba don ƙayyade yarjejeniya kashi (tabbatacce, korau, da gabaɗaya) da sauran sigogin kwatance don nazarin huɗun. Kowane kwatancen gwaji an yi shi da CRS, a cikin wannan binciken an saita CRS ta ka'ida "kowane tabbatacce" kuma an ƙayyade sakamakon, ba ta gwaji ɗaya ba, mun yi amfani da aƙalla sakamakon gwaji guda biyu da suka dace. Bugu da kari, game da watsa COVID-19, sakamakon mummunan sakamako na karya ya fi hatsari fiye da sakamakon karya. Don haka, don faɗi “tabbatacce” daidai gwargwadon sakamako daga sakamakon CRS, aƙalla gwaje-gwajen gwaji guda biyu dole ne su kasance tabbatacce, ma'ana cewa aƙalla sakamako mai kyau yana yiwuwa ya fito daga gwajin EUA. Don haka, daga cikin sakamakon gwaji guda huɗu, sakamakon gwaji biyu ko sama da haka waɗanda ke ba da sakamako iri ɗaya ana ɗaukarsu tabbatacce ne ko kuma mara kyau18,27.
An tattara bayanai ta hanyar amfani da sifofin cire bayanan da aka tsara, shigarwar bayanai da bincike an yi su ta amfani da software na ƙididdiga na Excel da sigar SPSS 23.0 don ƙididdige ƙididdiga. An bincika yarjejeniya mai inganci, mara kyau, da gabaɗaya, kuma an yi amfani da maki Kappa don tantance ƙimar yarjejeniya ta kowace hanya tare da CRS. Ana fassara ƙimar Kappa kamar haka: 0.01 zuwa 0.20 don yarjejeniya mai sauƙi, 0.21 zuwa 0.40 don yarjejeniya ta gaba ɗaya, 0.41-0.60 don yarjejeniya mai matsakaici, 0.61-0.80 don babbar yarjejeniya da 0.81-0.99 don cikakkiyar yarjejeniya28.
An sami izini na ɗabi'a daga Jami'ar Addis Ababa kuma duk ƙa'idodin gwaji don wannan binciken an amince da su daga Hukumar Kula da Da'a ta Kimiyya ta Cibiyar Kiwon Lafiyar Jama'a ta Habasha. Lambar tunani don Lasisin Da'a na EPHI shine EPHI/IRB-279-2020. An yi amfani da dukkan hanyoyin bisa ga shawarwari da tanadi na Ƙimar Jagorori na Ƙasar Habasha don Kula da COVID-19. Bugu da ƙari, an sami izinin rubutaccen bayani daga duk mahalarta binciken kafin shiga cikin binciken.
Duk bayanan da aka samu ko aka bincika a cikin wannan binciken an haɗa su a cikin wannan labarin da aka buga. Bayanai masu goyan bayan sakamakon wannan binciken suna samuwa daga marubucin bisa ga buƙatu mai ma'ana.
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