Imikorere ya acide nucleic aside amplification isobanura kumenya SARS-CoV-2 muri Etiyopiya

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Kuva indwara ya coronavirus yo mu 2019 (COVID-19) yatangira, ibizamini byinshi byo kongera aside nucleic acide (NAATs) byatejwe imbere kwisi yose kandi byabaye ibisubizo bisanzwe. Nubwo ibizamini byinshi byateguwe vuba kandi bigashyirwa mubizamini byo gusuzuma laboratoire, imikorere yibi bizamini ntabwo yasuzumwe muburyo butandukanye. Kubwibyo, ubu bushakashatsi bwari bugamije gusuzuma imikorere ya Abbott SARS-CoV-2, Daan Gene, BGI, na Sansure Biotech yerekana ikoresheje Composite Reference Standard (CRS). Ubushakashatsi bwakorewe mu kigo cy’ubuzima rusange cya Etiyopiya (EPHI) kuva ku ya 1 kugeza ku ya 30 Ukuboza 2020. Hakozwe 164 nasofaryngeal hakoreshejwe ibikoresho bito bya QIAamp RNA na sisitemu yo gutegura icyitegererezo cya ADN Abbott. Mubigereranyo 164, 59.1% byari byiza naho 40.9% byari bibi kuri CRS. Sansure Biotech positivité yari hasi cyane ugereranije na CRS (p <0.05). Sansure Biotech positivité yari hasi cyane ugereranije na CRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p <0,05). Ibisubizo byiza bya Sansure Biotech byari hasi cyane ugereranije na 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 yagize ibisubizo bike cyane ugereranije na CRS (p <0.05).Amasezerano rusange yisesengura ane yari 96.3–100% ugereranije na CRS. Usibye igipimo cyiza cya positivitike ya Sansure Biotech assay, imikorere yibisubizo bine byagereranijwe. Nkibyo, Sansure Biotech [Ubushakashatsi Gusa (RUO)] isaba bisaba izindi mpamyabushobozi kugirango ikoreshwe muri Etiyopiya. Hanyuma, ubushakashatsi bwinyongera bugomba gusuzumwa kugirango hasuzumwe ibisubizo hamwe nibisabwa nababikoze.
Kwipimisha muri laboratoire biri muri gahunda y’umuryango w’ubuzima ku isi (OMS) Gahunda y’ingamba zo kurwanya indwara ya Coronavirus 2019 (COVID-19) Kwitegura no gusubiza (SPRP). OMS itanga inama ko ibihugu bigomba kubaka ubushobozi bwa laboratoire kugira ngo biteze imbere imyiteguro, gucunga neza imanza, kuba maso no gukemura vuba ibibazo by’ubuzima rusange. Ibi byerekana ko uruhare rwa laboratoire ari urufunguzo rwo kuranga indwara na epidemiologiya y’indwara zandura zigaragara no kugenzura ikwirakwizwa ryazo.
Gupima COVID-19 bisaba amakuru ya epidemiologiya nubuvuzi, ibimenyetso / ibimenyetso byihariye, hamwe namakuru ya radiografiya na laboratoire2. Kuva icyorezo cya COVID-19 kivugwa i Wuhan mu Bushinwa, hakozwe ibizamini byinshi byo kongera aside nucleic acide (NAATs) ku isi. Igihe nyacyo cyo guhinduranya inyandiko ya polymerase yerekana (rRT-PCR) yakoreshejwe nkuburyo busanzwe kandi busanzwe bwo gusuzuma laboratoire ya syndrome ikaze yubuhumekero bukabije 2 (SARS-CoV-2) 3 kwandura. Kumenyekanisha molekulari ya SARS-CoV-2 mubusanzwe bishingiye kuri N (nucleocapsid protein gene), E (gene envelope protein), na RdRp (RNA iterwa na RNA polymerase gene) muri ORF1a / b (ifunguye ryo gusoma 1a / b) . gene) akarere kagaragaye kuri genome ya virusi. Bafatwa nk'uturere nyamukuru twabitswe dusanga muri virusi ya virusi kugirango tumenye virusi4. Muri ubwo bwoko, genes ya RdRp na E ifite sensibilité yo kumenya neza, naho N gen ifite sensibilité nkeya5.
Imikorere ya PCR isuzuma irashobora gutandukana bitewe nibintu bitandukanye nka: gukuramo reagent, amplification / detection reagents, uburyo bwo kuvoma, ubwiza bwimashini ya PCR nibindi bikoresho. Kugeza muri Mata 2020, ibikoresho birenga 48 bitandukanye byo gusuzuma byaturutse mu bihugu icyenda byabonye uruhushya rwo gukoresha byihutirwa (EUA) mu gusuzuma COVID-196. Muri Etiyopiya, porogaramu zirenga 14 nyazo za PCR zikoreshwa mu kumenya PCR kumenya SARS-CoV-2 ku bigo 26 by’ubuzima rusange, harimo ABI 7500, Abbott m2000, Roche 48000 na Quant-studio7. Byongeye kandi, ibikoresho bitandukanye bya PCR birahari, nk'ikizamini cya Daan Gene, ikizamini cya Abbott SARS-CoV-2, ikizamini cya Sansure Biotech, n'ikizamini cya SARS-CoV-2 BGI. Nubwo rRT-PCR yunvikana cyane, abarwayi bamwe na bamwe barwaye COVID-19 bavuga ingaruka mbi zitari nziza kubera kopi idahagije ya aside ya ribonucleic aside (RNA) mu byitegererezo kubera gukusanya nabi, gutwara, kubika no gufata neza, no gupima laboratoire. imiterere n'ibikorwa by'abakozi8. Byongeye kandi, icyitegererezo cyangwa kugenzura amakosa, gushiraho inzitizi (Ct), hamwe no guhuza imbaraga hamwe nandi acide ya nucleic acide cyangwa idakora / isigaye SARS-CoV-2 RNA irashobora gutuma habaho ibisubizo byiza mubisubizo bya rRT-PCR9. Rero, biragaragara ko ibizamini bya PCR bishobora rwose kwerekana abatwara ibice bya gene, kuko bidashobora no gutandukanya genes zikora virusi, bityo ibizamini birashobora kumenya gusa abatwara ntabwo ari abarwayi10. Kubwibyo, ni ngombwa gusuzuma imikorere yo gusuzuma ukoresheje uburyo busanzwe mugushiraho. Nubwo reagent nyinshi za NAAT ziraboneka mu kigo cy’ubuzima rusange cya Etiyopiya (EPHI) ndetse no mu gihugu hose, nta bisuzumabumenyi bigereranya byerekana imikorere yabyo. Kubwibyo, ubu bushakashatsi bwari bugamije gusuzuma imikorere igereranya ibikoresho biboneka mubucuruzi kugirango hamenyekane SARS-CoV-2 na rRT-PCR ukoresheje ingero zamavuriro.
Abitabiriye 164 bose hamwe bakekwaho kuba COVID-19 bashyizwe muri ubu bushakashatsi. Umubare munini w'icyitegererezo waturutse mu bigo nderabuzima (118/164 = 72%), mu gihe 46 (28%) basigaye bitabiriye ibigo bitavura. Mu bitabiriye amahugurwa batavuwe muri iki kigo, 15 (9.1%) bari bafite ibibazo by’amavuriro naho 31 (18.9%) bafite aho bahurira n’imanza zemejwe. Abitabiriye mirongo icyenda na batatu (56.7%) bari abagabo, naho imyaka (± SD) yabitabiriye yari 31.10 (± 11.82).
Muri ubu bushakashatsi, hagaragajwe ibipimo byiza n'ibibi by'ibizamini bine kuri COVID-19. Rero, igipimo cyiza cyibisubizo bya Abbott SARS-CoV-2, Daan Gene 2019-nCoV, SARS-CoV-2 BGI, na Sansure Biotech 2019-nCoV ni 59.1%, 58.5%, 57.9% na 55.5% . Amanota meza kandi meza yibintu (CRS) amanota 97 (59.1%) na 67 (40.9%), (Imbonerahamwe 1). Muri ubu bushakashatsi, ubusobanuro bwa CRS bwari bushingiye ku itegeko "ryiza ryose", aho mu bisubizo bine, ibisubizo bibiri cyangwa byinshi byatanze ibisubizo bimwe byafatwaga nkibyiza cyangwa bibi.
Muri ubu bushakashatsi, twasanze amasezerano mabi (NPA) ya 100% (95% CI 94,6–100) kubisesengura byose ugereranije na CRS. Isesengura ry’ibinyabuzima rya Sansure ryerekanye PPA ntoya ya 93.8% (95% CI 87.2-97.1) naho isesengura rya Daan Gene 2019-nCoV ryagize amasezerano rusange ya 99.4% (95% CI 96.6-99.9). Ibinyuranye, amasezerano rusange hagati ya SARS-CoV-2 BGI hamwe na Sansure Biotech 2019-nCoV yakozwe yari 98.8% na 96.3% (Imbonerahamwe 2).
Cohen coefficente ya kappa yamasezerano hagati ya CRS na Abbott SARS-CoV-2 ibisubizo byubushakashatsi byari bihuye neza (K = 1.00). Mu buryo nk'ubwo, indangagaciro za kappa za Cohen zagaragajwe na Daan Gene 2019-nCoV, SARS-CoV-2 BGI, na Sansure Biotech 2019-nCoV nazo zihuye neza na CRS (K ≥ 0.925). Muri iri sesengura ryagereranijwe, ikizamini cya chi-kare (ikizamini cya McNemar) cyerekanye ko ibisubizo bya Sansure Biotech 2019-nCoV ibisubizo bitandukanye cyane nibisubizo bya CRS (p = 0.031) (Imbonerahamwe 2).
Nkuko bigaragara ku gishushanyo.1 ijanisha ryagaciro ka Ct yo hasi (<20 Ct) ya Abbott SARS-CoV-2 assay (ihuriweho na RdRp na N gene) yari 87,6% naho ORF1a / b gene Ct agaciro ka Sansure Biotech 2019-nCoV yerekanaga ko ijanisha ryo hasi Agaciro Ct (<20 Ct) kari 50.3% naho Ct yo hejuru (36-40 Ct) yari 3.2%. 1 ijanisha ryagaciro ka Ct yo hasi (<20 Ct) ya Abbott SARS-CoV-2 assay (ihuriweho na RdRp na N gene) yari 87,6% naho ORF1a / b gene Ct agaciro ka Sansure Biotech 2019-nCoV yerekanaga ko ijanisha ryo hasi Agaciro Ct (<20 Ct) kari 50.3% naho Ct yo hejuru (36-40 Ct) yari 3.2%.Nkuko bigaragara ku gishushanyo.1. (<20 Ct) соверлял 50,3%, а высокое значение Ct (36-40 Ct) соверляло 3,2%. 1. ko ijanisha ryagaciro gake Ct (<20 Ct) ryagize 50.3%, naho agaciro gakomeye Ct (36-40 Ct) kangana na 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 7. 87,6%值 (<20 Ct) 的百分比为 50.3% ，高 Ct 值 (36-40 Ct) 的百分比为 3.2%。 Nkuko bigaragara ku gishushanyo 1, ijanisha ryo hasi ya Ct (<20 Ct) yikizamini cya Abbott SARS-CoV-2 (guhuza RdRp na N gene) ni 87,6%, ORF1a / b gene Ct agaciro ka Sansure Biotech 2019-nCoV yerekana Ct low (<20 Ct) 的 ijanisha ni 50.3%, 高 Ct 值 (36-40 Ct) 的 ijanisha ni 3.2%. Как показано на рисунке 1, мат Abbott SARS-CoV-2 (сочетающий зы RdRp и N) Imel самое низкое нанное значение Ct (<20 Ct) в размере 87,6%, а значение Ct чиа ORF1a / b в исследовании. - Аниза nCoV показал низкий Ct. Nkuko bigaragara ku gishushanyo 1, Abbott SARS-CoV-2 (guhuza genes ya RdRp na N) yari ifite ijanisha ryo hasi ya Ct agaciro (<20 Ct) kuri 87,6%, mugihe Ct agaciro ka ORF1a / b gene muri Sansure Ubushakashatsi bwa Biotech 2019 - Isesengura rya nCoV ryerekanye Ct nkeya. Процент значений (<20 Ct) соверил 50,3%, а в вококих значений Ct (36-40 Ct) совилил 3,2%. Ijanisha ryagaciro (<20 Ct) ryari 50.3%, naho ijanisha ryagaciro rya Ct (36-40 Ct) ryari 3.2%.Ikizamini cya Abbott SARS-CoV-2 B cyanditse Ct agaciro kari hejuru ya 30. Ku rundi ruhande, kuri BGI SARS-CoV-2 assay ORF1a / b gene yari ifite agaciro gakomeye ka Ct (> 36 Ct) ijanisha rya 4% (Ishusho 1). Ku rundi ruhande, kuri BGI SARS-CoV-2 assay ORF1a / b gene yari ifite agaciro gakomeye ka Ct (> 36 Ct) ijanisha rya 4% (Ishusho 1). С. Ku rundi ruhande, mu isesengura rya BGI SARS-CoV-2 gene ORF1a / b yari ifite agaciro gakomeye Ct (> 36 Ct), ijanisha ryayo rikaba 4% (Ishusho 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 ）。 Ku rundi ruhande, muri BGI SARS-CoV-2 gutahura, ijanisha rya ORF1a / b gene ifite agaciro ka Ct (> 36 Ct) ni 4% (Ishusho 1). С. Ku rundi ruhande, mu isesengura rya BGI SARS-CoV-2, ijanisha rya gen ORF1a / b rifite agaciro gakomeye ka Ct (> 36 Ct) ryari 4% (Ishusho 1).
Muri ubu bushakashatsi, twafashe ibyitegererezo 164 bya nasofaryngeal. Kubwoko bwose bwibisubizo, RNA kwigunga no kongera imbaraga byakozwe hakoreshejwe uburyo nibikoresho byasabwe nababikora.
Ubu bushakashatsi bwerekanye ko ikizamini cya Abbott kuri SARS-CoV-2 gifite imikorere imwe yo kumenya nka CRS, hamwe 100% byiza, bibi, kandi muri rusange. Amasezerano ya kappa ya Cohen ni 1.00, yerekana amasezerano yuzuye na CRS. Ubushakashatsi nk'ubwo bwakozwe na kaminuza ya Washington muri Amerika bwerekanye ko muri rusange ubushakashatsi bwakozwe na Abbott kuri SARS-CoV-2 bwari 93% na 100%, ugereranije n'ubushakashatsi bwakozwe na laboratoire (LDA) bwa CDC . 11. Sisitemu yo gutahura Abbott SARS-CoV-2 ishingiye ku kumenya icyarimwe icyarimwe cyo kumenya gen na N na RdRp, kubera ko genes zombi zumva cyane, zigabanya ibibi 12. Ubushakashatsi bwakorewe i Vienne muri Otirishiya bwerekanye kandi ko ingano nini yo gukuramo urugero hamwe n’ubunini bwa eluent byagaragaye byagabanije ingaruka zo kugabanuka no kongera ubushobozi bwo gutahura13. Rero, Abbott ihuye neza na SARS-CoV-2 isuzuma irashobora guhuzwa na sisitemu yo gutahura icyarimwe icyarimwe ikamenya genes ikomatanya, ikuramo umubare munini wintangarugero (0,5 ml), kandi ikoresha urugero rwinshi (40 µl).
Ibisubizo byacu byerekanaga kandi ko imikorere yo kumenya ikizamini cya genetike ya Daan yari imeze nka CRS. Ibi bihuye nubushakashatsi14 bwakorewe muri kaminuza ya Anhui i Huainan, mu Bushinwa, ndetse n’uko uruganda rusaba amasezerano 100%. Nubwo amakuru y’ibisubizo bihamye, icyitegererezo kimwe cyari kibi nyuma yo kongera kugerageza umwe, ariko cyabaye cyiza muri Abbott SARS-CoV-2 na Sansure Biotech nCoV-2019. Ibi birerekana ko hashobora kubaho impinduka mubisubizo muburyo butandukanye bwo gusuzuma. Nubwo bimeze bityo ariko, mubushakashatsi bwakorewe mubushinwa15, ibisubizo byubushakashatsi bwa Daan Gene byari bitandukanye cyane (p <0.05) ugereranije na laboratoire yabo yasobanuwe. Nubwo bimeze bityo ariko, mubushakashatsi bwakorewe mubushinwa15, ibisubizo byubushakashatsi bwa Daan Gene byari bitandukanye cyane (p <0.05) ugereranije na laboratoire yabo yasobanuwe. Тем не мене, в исследовании, проведенном в Китае15, результат мата Daan Gene значительно отличался (p <0,05) от ла лабораторного э эононного мата. Nyamara, mu bushakashatsi bwakorewe mu Bushinwa15, ibisubizo by'isesengura rya Daan Gene byari bitandukanye cyane (p <0.05) n'isesengura ryabo rya laboratoire.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差异（ 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相比有显着差 <0.05 Однако в исследовании, проведенном в Китае15, результаты зетического теста Daan значительно отличались (p <0,05) по сравнению с его э эононным лабориторным чомом. Nyamara, mu bushakashatsi bwakorewe mu Bushinwa15, ibyavuye mu kizamini cya geneti Daan byari bitandukanye cyane (p <0.05) ugereranije n’ikizamini cya laboratoire.Uku kunyuranya gushobora guterwa no kumva neza ikizamini cyerekeranye no kumenya SARS-CoV-2, kandi ubushakashatsi burashobora kuba ingenzi kugirango umenye icyabiteye.
Byongeye kandi, ubushakashatsi bwacu bwasuzumye imikorere igereranya ya SARS-CoV-2 BGI hamwe na CRS, yerekana amasezerano meza meza (PPA = 97.9%), amasezerano mabi (NPA = 100%), hamwe namasezerano rusange yuburinganire (uburinganire () OPA). ). = 98.8%). Indangagaciro za Kappa ya Cohen yerekanye amasezerano meza (K = 0.975). Ubushakashatsi mu Buholandi16 n'Ubushinwa15 bwerekanye ibisubizo bihamye. Ikizamini cya SARS-CoV-2 BGI ni gene imwe (ORF1a / b) ikizamini cyo kumenya ukoresheje 10 µl amplification / detection eluate. Nubwo amasezerano meza y'ibarurishamibare hamwe n'ibisubizo byatanzwe, isesengura ryabuze ingero ebyiri nziza (1.22%) z'icyitegererezo cyose. Ibi birashobora kugira ingaruka zikomeye kumavuriro yanduye haba kumurwayi ndetse no kurwego rwabaturage.
Ubundi isesengura rigereranya ryashyizwe muri ubu bushakashatsi ni Sansure Biotech nCoV-2019 rRT-PCR (RUO); ijanisha rusange ryimikino ryari 96.3%. Imbaraga z’amasezerano zagenwe kandi n’agaciro ka Kahen ya Cohen, yari 0.925, byerekana amasezerano yuzuye na CRS. Na none kandi, ibisubizo byacu birasa n’ubushakashatsi bwakorewe muri kaminuza yo mu majyepfo ya Changsha, mu Bushinwa, no mu ishami rya Laboratwari ishami ry’ibitaro by’abaturage bya Liuzhou, Umujyi wa Liuzhou, mu Bushinwa17. Nubwo ibivugwa mu mibare byavuzwe haruguru byanditswe, ikizamini cya chi-kare (ikizamini cya MacNemar) cyerekanye ko ibisubizo by’ubushakashatsi bwa Sansure Biotech byagize itandukaniro rishingiye ku mibare ugereranije na CRS (p <0.005). Nubwo ibivugwa mu mibare byavuzwe haruguru byanditswe, ikizamini cya chi-kare (ikizamini cya MacNemar) cyerekanye ko ibisubizo by’ubushakashatsi bwa Sansure Biotech byagize itandukaniro rishingiye ku mibare ugereranije na CRS (p <0.005). Несмотря на то, что было зафик Кановано укаزанное вше хорошее статистическое соответствие, критерий хи-бар (критерий Макнемара) показал, что результат мата Sansure Biotech имеет стачи скичи <0,005). Nubwo amasezerano meza y’ibarurishamibare yavuzwe haruguru yanditswe, ikizamini cya chi-kare (ikizamini cya McNemar) cyerekanye ko ibisubizo by’ubushakashatsi bwa Sansure Biotech bifite itandukaniro rinini cyane ugereranije na 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 检测的结果与 CRS 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 n n （（ macnemar 检验 表明 ， ， sansure biotech 检测 结果 与 crs 相比 具有 显着 （（ p <0.005 。。。。。。。。。。。。。。。。 。。。）））） Несмотря на отмеченное вше хорошее статистическое соответствие, критерий хи-нан (критерий Макнемара) показал статистически значимую разницу (p <0,005) межудуом Sansure Biotech и CRS. Nubwo amasezerano meza y'ibarurishamibare yavuzwe haruguru, ikizamini cya chi-kare (ikizamini cya McNemar) cyerekanye itandukaniro rinini cyane (p <0.005) hagati ya Sansure Biotech assay na CRS.Ingero esheshatu (3,66%) wasangaga ari ibinyoma ugereranije na CRS (Imbonerahamwe y'inyongera 1); ibi ni ngombwa cyane, cyane cyane urebye imbaraga zo kwanduza virusi. Amakuru yavuzwe haruguru nayo ashyigikira iki gipimo gito cyo gutahura15.
Muri ubu bushakashatsi, indangagaciro za Ct zagenwe kuri buri cyegeranyo hamwe na platifomu bijyanye, hamwe n’igiciro cyo hasi ya Ct ivugwa muri Abbott SARS-CoV-2. Ibisubizo birashobora kuba bifitanye isano na Abbott icyarimwe sisitemu yo gupima genetike yo kumenya SARS-CoV-2. Kubwibyo, ukurikije Ishusho ya 1, 87,6% y ibisubizo bya Abbott SARS-CoV-2 byari bifite Ct agaciro kari munsi ya 20. Gusa umubare muto wibisubizo (12.4%) byari murwego rwa 20-30. Indangagaciro za Ct ziri hejuru ya 30 ntabwo zanditswe. Usibye kuba Abbott yarakoresheje uburyo bwo gupima genetike ya SARS-CoV-2, iki gisubizo gishobora kuba gifitanye isano n’umupaka wo hasi (32.5 RNA kopi / mL) 18, ibyo bikaba bikubye inshuro eshatu ugereranije n’urwego rwo hasi rw’isosiyete 100 kopi ya RNA; / mL. ml) 19.
Ubu bushakashatsi bufite aho bugarukira: icya mbere, ntabwo dufite uburyo busanzwe / bwerekana [nk'umutwaro wa virusi cyangwa ibindi bizamini bya laboratoire (LDA)] kubera kubura amikoro. Icya kabiri, ingero zose zikoreshwa muri ubu bushakashatsi zari izuru rya nasofaryngeal, mugihe ibisubizo bitakurikizwa kubundi bwoko bw'icyitegererezo, naho icya gatatu, ingano y'icyitegererezo yari nto.
Ubu bushakashatsi bwagereranije imikorere ya rRT-PCR isuzuma SARS-CoV-2 ukoresheje ingero za nasofaryngeal. Ubushakashatsi bwose bwakozwe bwagize imikorere igereranywa, usibye ibizamini bya Sansure Biotech. Uretse ibyo, igipimo cyiza cyagaragaye mu bushakashatsi bwa Sansure Biotech ugereranije na CRS (p <0.05). Uretse ibyo, igipimo cyiza cyagaragaye mu bushakashatsi bwa Sansure Biotech ugereranije na CRS (p <0.05). Кроме того, в Тее Sansure Biotech был вызвлен низкий пол положительных результатов по сравнению с CRS (p <0,05). Mubyongeyeho, ikizamini cya Sansure Biotech cyerekanye ijanisha rito ryibisubizo byiza ugereranije na 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, мат Sansure Biotech imel boлее низкий оровень положительных результатов по сравнению с CRS (p <0,05). Byongeye kandi, Sansure Biotech assay yari ifite igipimo cyiza cyo hasi ugereranije na CRS (p <0.05).Isesengura rya Sansure Biotech nCoV-2019 (RUO) isesengura rya PPA, NPA n'amasezerano rusange yarenze 93.5% hamwe na Cohen Kappa imbaraga zamasezerano zifite agaciro ka 0.925. Hanyuma, Sansure Biotech Assay (RUO) ikeneye kwemezwa kugirango ikoreshwe muri Etiyopiya, kandi ubushakashatsi bwiyongereye bugomba gusuzumwa kugirango harebwe ibisabwa n'ababikora ku giti cyabo.
Igishushanyo mbonera cyagereranijwe cyakorewe ku bigo nderabuzima bine biri i Addis Abeba, mu bitaro bya Eka Kotebe, mu kigo cyita ku itorero rya Millennium, mu bitaro by’Urwibutso rwa Zewooditu, no mu bitaro by’inzobere by’igituntu cya Mutagatifu Petero. Amakuru yakusanyijwe hagati yitariki ya 1 na 31 Ukuboza 2020. Ibigo nderabuzima by’ubu bushakashatsi byatoranijwe nkana bitewe n’imibare yabo myinshi ndetse n’ibigo nderabuzima bikomeye byo mu mujyi. Mu buryo nk'ubwo, ibikoresho, birimo ABI 7500 na Abbott m2000 ibikoresho nyabyo bya PCR, byatoranijwe hakurikijwe ibyifuzo by’abakora reagent ya NAAT, kandi hatoranijwe ibikoresho bine bya PCR byatoranijwe muri ubu bushakashatsi, kubera ko laboratoire nyinshi zo muri Etiyopiya zakoresheje byibuze bane muri bo. Ikizamini cya Gene, Abbott SARS-CoV-2 ikizamini, Ikizamini cya Sansure Biotech, na SARS-CoV-2 BGI ikizamini cyakozwe mugihe cyo kwiga).
Ikizamini cya SARS-CoV-2 cyakozwe kuva ku ya 1 kugeza ku ya 30 Ukuboza 2020 hifashishijwe ml 3 ya Virusi yo Gutwara Virusi (VTM) (Ikoranabuhanga rya Miraclean, Shenzhen, Ubushinwa) ku bantu barimo gukorerwa iperereza kuri COVID-19 ivugwa kuri EPHI. Urugero rwa Nasopharyngeal rwakusanyirijwe hamwe nabakusanyije icyitegererezo bazoherezwa muri EPHI mumapaki atatu. Mbere yo gutandukanya aside nucleic, buri sample ihabwa numero yihariye iranga. Gukuramo bikorwa muri buri cyitegererezo ako kanya ukihagera ukoresheje uburyo bwo gukuramo intoki. Rero, kugirango ikurwe ryikora rya Abbott m2000, miriyoni 1,3 (harimo na 0.8 ml yapfuye nubunini bwa 0,5 ml yo gukuramo inlet) yikitegererezo yakuwe muri buri cyitegererezo hanyuma inyuzwa muri sisitemu yo gutegura ADN ya Abbott (Abbott Molecular Inc. des Plaines, IL, Amerika). ) Icyiciro cya 96 [92 by'icyitegererezo, kugenzura bibiri kugenzura no kugenzura bibiri bitari inyandikorugero (NTC)] byashyizwe mubikorwa rusange (kugarura no gutahura) ibyiciro bibiri bya SARS-CoV-2 (EUA) mugihe nyacyo. ubucukuzi bw'amabuye y'agaciro. Muri ubwo buryo ,, kubikuramo intoki, koresha ingero zimwe (kubikuramo byikora no kuvumbura). Niyo mpamvu, muri gahunda zose, 140 µl ntangarugero zarakuweho kandi zavomwe hakoreshejwe QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Ubudage) mubice 24 (harimo ingero 20, kugenzura bibiri hamwe na NTC ebyiri) mubice icyenda. Intoki zavanywemo intoki zongerewe kandi zimenyekana hifashishijwe igare ry’umuriro ABI 7500 ukoresheje SARS-CoV-2 BGI, Daan Gene assay, hamwe na Sansure Biotech.
Kwigunga byikora no kweza SARS-CoV-2 virusi RNA ikurikiza ihame ryamasaro ya magneti ikoresheje reagent ya Abbott ADN. Kudakora ingero no gukemura ibice bya virusi bikorwa hakoreshejwe icyuma kirimo isuku ya guanidine isothiocyanate kugirango yamagane poroteyine kandi idakora RNase. RNA noneho itandukanijwe na poroteyine no gutandukanya icyiciro gikomeye ukoresheje silika, ni ukuvuga umunyu wa guanidinium na alkaline pH ya buffer ya lysis itera guhuza aside nucleic na silika (SiO2). Intambwe yo koza ikuraho poroteyine zisigaye hamwe n’imyanda kugirango bitange igisubizo kiboneye. RNA iboneye itandukanijwe na microparticles ishingiye kuri silika ikoresheje imbaraga za magnetiki igikoresho 20,21. Kurundi ruhande, kwigunga intoki no kweza RNA bikorwa nuburyo bwa spin inkingi hakoreshejwe centrifugation aho guhagarara magnetique no gutandukanya microparticles na eluent.
Ikizamini cya Abbott Real-Time SARS-CoV-2 Ikizamini cyo Kumenya (Abbott Molecular, Inc.) cyakozwe hakurikijwe amabwiriza yabakozwe, yakiriye EUA 19,22 na OMS na FDA. Muri iyi protocole, icyitegererezo cyo kudakora mbere yo gukuramo cyakorewe mu bwogero bwamazi kuri 56 ° C kuminota 30. Nyuma yo kudakora virusi, gukuramo aside nucleic byakorewe ku gikoresho cya Abbott m2000 SP kuva kuri 0,5 ml VTM hakoreshejwe uburyo bwo gutegura ADN ya Abbott m2000. nk'uko uwabikoze abitangaza. Amplification no gutahura byakozwe hakoreshejwe igikoresho cya Abbott m2000 RT-PCR, naho gutahura kabiri byakorewe genes ya RdRp na N. ROX) na VIC P (irangi ryihariye) mugushaka no kumenya kugenzura imbere, kwemerera icyarimwe kumenya ibicuruzwa byombi byongera 19.
Uburyo bwa amplification detection yiki gikoresho bushingiye ku ntambwe imwe ya tekinoroji ya RT-PCR. Ingirabuzimafatizo za ORF1a / b na N zatoranijwe nk'uturere twabungabunzwe na Daan Gene Technology kugira ngo tumenye akarere kongerewe imbaraga. Primers yihariye na fluorescente (N gene probe yanditseho FAM, ORF1a / b probe yanditseho VIC) yateguwe kugirango tumenye SARS-CoV-2 RNA mubitegererezo. Amaherezo ya eluent na master mixes yateguwe hongerwaho 5 µl ya eluent kuri 20 µl ya master mix ivanze nubunini bwa 25 µl. Amplification and detection byakorewe icyarimwe kubikoresho bya ABI 750024-nyabyo-PCR.
Ingirabuzima fatizo za ORF1a / b na N zagaragaye hakoreshejwe Sansure Biotech nCoV-2019 Nucleic Acide Diagnostic Kit (fluorescent PCR detection). Tegura iperereza ryihariye kuri buri gen igamije guhitamo umuyoboro wa FAM mukarere ka ORF1a / b hamwe numuyoboro wa ROX kuri N. Kuri iki gikoresho cyo gusuzuma, eluent na master mix reagents yongeweho kuburyo bukurikira: tegura 30 µl master mix reagent na 20 µl eluted sample yo gutahura / amplification. Igihe nyacyo PCR ABI 750025 yakoreshejwe muri amplification / detection.
Ikizamini cya SARS-CoV-2 BGI ni fluorescent nyayo-rRT-PCR ibikoresho byo gusuzuma COVID-19. Agace kagenewe kari mu karere ka ORF1a / b ka genoside ya SARS-CoV-2, nuburyo bumwe bwo kumenya gene. Byongeye kandi, gene yo mu rugo yumuntu act-actin ni gen igenwa imbere. Igishushanyo mbonera cyateguwe mukuvanga 20 µl ya master mix reagent na 10 µl yicyitegererezo cya RNA cyakuwe mubisahani26. ABI 7500 fluorescent numero nyayo-igikoresho cya PCR yakoreshejwe mugukomeza no gutahura. Kwiyongera kwa nucleic aside yose, PCR ikora kuri buri cyegeranyo, no gusobanura ibisubizo byakozwe ukurikije amabwiriza yabakozwe (Imbonerahamwe 3).
Muri iri sesengura rigereranya, ntabwo twakoresheje uburyo busanzwe bwo gukoresha kugirango tumenye amasezerano ku ijana (ibyiza, ibibi, na rusange) nibindi bipimo byo kugereranya kubisesenguye bine. Kugereranya ibizamini byose byakorewe hamwe na CRS, muri ubu bushakashatsi CRS yashyizweho n amategeko "icyiza cyose" kandi ibisubizo byagenwe, ntabwo byakozwe nikizamini kimwe, twakoresheje byibuze ibisubizo bibiri bihuye. Mubyongeyeho, kubijyanye no kwanduza COVID-19, ibisubizo bibi bibi ni bibi cyane kuruta ibisubizo byiza. Kubwibyo, kuvuga "positif" uko bishoboka kwose bivuye mubisubizo bya CRS, byibuze ibizamini bibiri byibisubizo bigomba kuba byiza, bivuze ko byibuze igisubizo kimwe cyiza gishobora guturuka mubibazo bya EUA. Rero, mubisubizo bine byikizamini, bibiri cyangwa byinshi ibisubizo byikizamini bitanga ibisubizo bimwe bifatwa nkibyiza cyangwa bibi 18,27.
Amakuru yakusanyijwe hifashishijwe impapuro zo gukuramo amakuru yubatswe, kwinjiza amakuru no gusesengura byakozwe hifashishijwe porogaramu ya Excel y'ibarurishamibare na verisiyo ya SPSS 23.0 ku mibare isobanura. Hasesenguwe amasezerano meza, mabi, kandi muri rusange ku ijana, kandi amanota ya Kappa yakoreshejwe kugirango hamenyekane urwego rwamasezerano ya buri buryo na CRS. Indangagaciro za Kappa zisobanurwa gutya: 0.01 kugeza 0.20 kubwumvikane bworoheje, 0.21 kugeza 0.40 kumasezerano rusange, 0.41-0.60 kubwumvikane buciriritse, 0.61-0.80 kubwamasezerano akomeye na 0.81-0.99 kubwumvikane bwuzuye28.
Imyitwarire myiza yabonetse muri kaminuza ya Addis Abeba kandi protocole zose zigeragezwa kuri ubu bushakashatsi zemejwe n’ikigo cy’ubushakashatsi bw’imyitwarire y’ubumenyi bw’ikigo cya Etiyopiya. Inomero yerekana uruhushya rwimyitwarire ya EPHI ni EPHI / IRB-279-2020. Uburyo bwose bwakoreshejwe hakurikijwe ibyifuzo n’ingingo z’amabwiriza rusange y’igihugu cya Etiyopiya yo kuvura COVID-19. Byongeye kandi, inyandiko yanditse yabimenyeshejwe yabonetse mubitabiriye kwiga mbere yo kwitabira kwiga.
Amakuru yose yabonetse cyangwa yasesenguwe muri ubu bushakashatsi yashyizwe muri iyi ngingo yatangajwe. Amakuru ashyigikira ibisubizo byubu bushakashatsi araboneka kubwanditsi abisabwe neza.
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