Ts'ebetso ea liteko tse 'ne tsa nucleic acid amplification ho khetholla SARS-CoV-2 naheng ea Ethiopia

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Ho tloha ha ho qhoma lefu la coronavirus (COVID-19) la 2019, liteko tse ngata tsa khoebo tsa nucleic acid amplification (NAATs) li entsoe lefatšeng ka bophara 'me li fetohile liteko tse tloaelehileng. Le hoja liteko tse 'maloa li ile tsa hlahisoa ka potlako' me tsa sebelisoa litekong tsa tlhahlobo ea laboratori, ts'ebetso ea liteko tsena ha e e-s'o hlahlojoe maemong a sa tšoaneng. Ka hona, boithuto bona bo ne bo reretsoe ho lekola ts'ebetso ea Abbott SARS-CoV-2, Daan Gene, BGI, le Sansure Biotech assays ho sebelisa Composite Reference Standard (CRS). Thuto e ile ea etsoa Setsing sa Bophelo bo Botle ba Sechaba sa Ethiopia (EPHI) ho tloha 1 ho ea ho 30 December 2020. Li-sampuli tsa 164 tsa nasopharyngeal li ile tsa ntšoa ka ho sebelisa QIAamp RNA mini kit le mokhoa oa ho lokisetsa sampole ea Abbott DNA. Har'a mehlala e 164, 59.1% e ne e le ntle 'me 40.9% e ne e le mpe bakeng sa CRS. Sansure Biotech positivity e ne e le tlase haholo ha e bapisoa le CRS (p <0.05). Sansure Biotech positivity e ne e le tlase haholo ha e bapisoa le CRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p <0,05). Liphetho tse ntle tsa Sansure Biotech li ne li le tlase haholo ha li bapisoa le 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 e bile le liphetho tse ntle tse fokolang haholo ha li bapisoa le CRS (p <0.05).Tumellano e akaretsang ea litlhahlobo tse 'ne e ne e le 96.3-100% ha e bapisoa le CRS. Ntle le sekhahla se tlase sa positivity ea tlhahlobo ea Sansure Biotech, ts'ebetso ea litlhahlobo tse 'ne e ne e batla e bapisoa. Ka hona, tlhahlobo ea Sansure Biotech [Research Only (RUO)] e hloka netefatso e eketsehileng bakeng sa tšebeliso ea eona Ethiopia. Qetellong, lipatlisiso tsa tlatsetso li lokela ho nahanoa ho lekola liteko ka likopo tse nepahetseng tsa moetsi.
Teko ea laboratori ke karolo ea Leano la Leano la Mokhatlo oa Lefatše oa Bophelo (WHO) bakeng sa Mafu a Coronavirus a 2019 (COVID-19) Ho Itokisa le Karabelo (SPRP). WHO e eletsa hore linaha li hloka ho haha ​​​​bokhoni ba laboratori ho ntlafatsa boitokiso, tsamaiso e nepahetseng ea linyeoe, ho falimeha le ho arabela ka potlako ho mathata a bophelo bo botle ba sechaba. Sena se fana ka maikutlo a hore karolo ea laboratori ke senotlolo sa ho khetholla lefu le lefu la seoa la likokoana-hloko tse hlahang le ho laola ho ata ha tsona.
Tlhahlobo ea COVID-19 e hloka lintlha tsa lefu la seoa le tsa bongaka, matšoao/matšoao a botho, le lintlha tsa radiographic le laboratori2. Ho tloha ha koluoa ​​​​ea COVID-19 e tlalehoa Wuhan, China, liteko tse ngata tsa khoebo tsa nucleic acid amplification (NAATs) li entsoe lefatšeng ka bophara. Real-time reverse transcription polymerase chain reaction (rRT-PCR) e sebelisitsoe e le mokhoa o tloaelehileng oa tlhahlobo ea laboratori ea ts'oaetso e matla ea ho hema ea 2 (SARS-CoV-2)3. Ho lemoha ha limolek'hule tsa SARS-CoV-2 hangata ho ipapisitse le N (nucleocapsid protein gene), E (enfelopo protheine gene), le RdRp (RNA-dependent RNA polymerase gene) ho ORF1a/b (foreimi e bulehileng ea ho bala 1a/b) . gene) sebaka se khethiloeng ho tsoa ho kokoana ea lefutso. Li nkoa e le libaka tse ka sehloohong tse sirelelitsoeng tse fumanoang ho liphatsa tsa lefutso tsa kokoana-hloko bakeng sa ho lemoha kokoana-hloko4. Har'a liphatsa tsa lefutso tsena, liphatsa tsa lefutso tsa RdRp le E li na le kutlo e phahameng ea tlhahlobo ea tlhahlobo, ha mofuta oa N o na le kutlo e tlase ea tlhahlobo5.
Ts'ebetso ea litlhahlobo tsa PCR e ka fapana ho latela lintlha tse fapaneng tse kang: li-reagents tsa ho hula, li-amplification / detection reagents, mokhoa oa ho ntša, boleng ba mochine oa PCR le lisebelisoa tse ling. Ho tloha ka Mmesa 2020, lisebelisoa tse fetang 48 tse fapaneng tsa tlhahlobo ea mafu tse tsoang linaheng tse robong li amohetse Authorization ea Tšebeliso ea Tšohanyetso (EUA) bakeng sa tlhahlobo ea COVID-196. Naheng ea Ethiopia, ho sebelisoa li-PCR tse fetang 14 tsa nako ea sebele bakeng sa ho lemoha SARS-CoV-2 litsing tsa bophelo tsa sechaba tse 26, ho kenyeletsoa ABI 7500, Abbott m2000, Roche 48000 le Quant-studio7. Ntle le moo, lisebelisoa tse fapaneng tsa liteko tsa PCR lia fumaneha, joalo ka tlhahlobo ea Daan Gene, tlhahlobo ea Abbott SARS-CoV-2, tlhahlobo ea Sansure Biotech, le tlhahlobo ea SARS-CoV-2 BGI. Leha rRT-PCR e na le kutloelo-bohloko haholo, bakuli ba bang ba nang le COVID-19 ba tlaleha liphetho tse mpe tsa bohata ka lebaka la ho haella ha likopi tsa viral ribonucleic acid (RNA) lisampole ka lebaka la pokello e sa nepahalang, lipalangoang, polokelo le ho ts'oaroa, le liteko tsa laboratori. maemo le diketso tsa basebetsi8. Ntle le moo, sampole kapa ho laola ho se sebetse hantle, tlhophiso ea li-cycle threshold (Ct), le ho kopana hape le li-nucleic acid tse ling tsa pathogenic kapa tse sa sebetseng / tse setseng tsa SARS-CoV-2 RNA li ka lebisa ho litholoana tse ntle tse fosahetseng litekong tsa rRT-PCR9. Kahoo, ho hlakile hore liteko tsa PCR ka sebele li ka khetholla bajari ba liphatsa tsa lefutso, kaha ba sitoa le ho khetholla pakeng tsa liphatsa tsa lefutso tse sebetsang e le kannete, kahoo liteko li ka khetholla feela bajari eseng bakuli10. Ka hona, ho bohlokoa ho lekola ts'ebetso ea tlhahlobo ho sebelisa mekhoa e tloaelehileng maemong a rona. Leha li-reagents tse ngata tsa NAAT li fumaneha Setsing sa Bophelo bo Botle sa Sechaba sa Ethiopia (EPHI) le naheng ka bophara, ha ho na tlhahlobo ea papiso ea katleho ea tsona e seng e tlalehiloe. Ka hona, boithuto bona bo ne bo reretsoe ho lekola ts'ebetso ea papiso ea lisebelisoa tse fumanehang khoebong bakeng sa ho fumanoa ha SARS-CoV-2 ke rRT-PCR ho sebelisoa mehlala ea bongaka.
Kakaretso ea bankakarolo ba 164 ba belaelloang hore ke COVID-19 ba kenyelelitsoe boithutong bona. Bongata ba lisampole bo ne bo tsoa litsing tsa kalafo (118/164 = 72%), athe barupeluoa ba setseng ba 46 (28%) ba ne ba tsoa litsing tseo e seng tsa kalafo. Har'a barupeluoa ba sa kang ba phekoloa setsing, ba 15 (9.1%) ba ne ba e-na le linyeoe tse belaelloang 'me ba 31 (18.9%) ba ne ba e-na le mabitso a linyeoe tse tiisitsoeng. Barupeluoa ba mashome a robong a metso e meraro (56.7%) e ne e le banna, 'me lilemo tse bolelang (± SD) tsa barupeluoa e ne e le lilemo tse 31.10 (± 11.82).
Phuputsong ena, litekanyetso tse ntle le tse mpe tsa liteko tse 'ne tsa COVID-19 li ile tsa khethoa. Kahoo, litefiso tse ntle tsa tlhahlobo ea Abbott SARS-CoV-2, Daan Gene 2019-nCoV assay, SARS-CoV-2 BGI assay, le tlhahlobo ea Sansure Biotech 2019-nCoV e ne e le 59.1%, 58.5%, 57.9% le 55.5% ka ho latelana. . Lintlha tse ntle le tse mpe tsa composite reference standard (CRS) e ne e le 97 (59.1%) le 67 (40.9%), ka ho latellana (Letlapa la 1). Phuputsong ena, tlhaloso ea CRS e ne e ipapisitse le molao oa "phoso efe kapa efe", moo ho liphetho tse 'ne tsa liteko, liphetho tse peli kapa ho feta tse faneng ka sephetho se ts'oanang li nkuoe li le ntle kapa li le mpe.
Thutong ena, re fumane tumellano ea liphesente tse mpe (NPA) ea 100% (95% CI 94.6-100) bakeng sa litlhahlobo tsohle ha li bapisoa le CRS. Tlhahlobo ea Sansure Biotechnology e bonts'itse PPA e nyane ea 93.8% (95% CI 87.2-97.1) mme tlhahlobo ea Daan Gene 2019-nCoV e bile le tumellano ea kakaretso ea 99.4% (95% CI 96.6-99.9). Ka lehlakoreng le leng, tumellano e akaretsang lipakeng tsa tlhahlobo ea SARS-CoV-2 BGI le tlhahlobo ea Sansure Biotech 2019-nCoV e ne e le 98.8% le 96.3%, ka ho latellana (Letlapa la 2).
Cohen's kappa coefficient ea tumellano pakeng tsa liphetho tsa tlhahlobo ea CRS le Abbott SARS-CoV-2 e ne e lumellana ka botlalo (K = 1.00). Ka mokhoa o ts'oanang, litekanyetso tsa Cohen's kappa tse fumanoeng ke Daan Gene 2019-nCoV, SARS-CoV-2 BGI, le Sansure Biotech 2019-nCoV le tsona li lumellana ka botlalo le CRS (K ≥ 0.925). Tlhahlobong ena ea papiso, tlhahlobo ea chi-square (teko ea McNemar) e bonts'itse hore liphetho tsa tlhahlobo ea Sansure Biotech 2019-nCoV li ne li fapane haholo le liphetho tsa CRS (p = 0.031) (Letlapa la 2).
Joalokaha ho bontšitsoe ho Feiga.1 peresente ea boleng bo tlase ba Ct (< 20 Ct) ea Abbott SARS-CoV-2 assay (e kopantseng RdRp le N gene) e ne e le 87.6% le boleng ba Ct ea mofuta oa ORF1a/b ea Sansure Biotech 2019-nCoV e bonts'itse hore peresente ea tlase Ct boleng (<20 Ct) e ne e le 50.3% le boleng bo phahameng ba Ct (36–40 Ct) e ne e le 3.2%. 1 peresente ea boleng bo tlase ba Ct (< 20 Ct) ea Abbott SARS-CoV-2 assay (e kopantseng RdRp le N gene) e ne e le 87.6% le boleng ba Ct ea mofuta oa ORF1a/b ea Sansure Biotech 2019-nCoV e bonts'itse hore peresente ea tlase Ct boleng (<20 Ct) e ne e le 50.3% le boleng bo phahameng ba Ct (36–40 Ct) e ne e le 3.2%.Joalokaha ho bontšitsoe ho Feiga.1, процент наименьшего значения Ct (< 20 Ct) анализа Abbott SARS-CoV-2 (комбинированный ген RdRp и N) составил 87,6%, а знагзаена Санкеение ORF1 2019-nCoV показало что процент низкого значения Ct (< 20 Ct) составлял 50,3%, а высокое значение Ct (36–40 Ct) составляло 3,2%. 1, peresente ea boleng bo tlase ka ho fetesisa ba Ct (<20 Ct) tlhahlobo ea Abbott SARS-CoV-2 (e kopantsoeng ea mofuta oa RdRp le N) e ne e le 87.6%, 'me boleng ba Ct ba tlhahlobo ea mofuta oa ORF1a/b ea Sansure Biotech 2019-nCoV e bontšitse. hore peresente ea boleng bo tlase ba Ct (<20 Ct) e ne e le 50.3%, le boleng bo phahameng ba Ct. (36–40 Ct) e ne e le 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1sure9-CoV2%, Sankade87.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. Joalokaha ho bontšitsoe setšoantšong sa 1, peresente e tlase ea boleng ba Ct (<20 Ct) ea tlhahlobo ea Abbott SARS-CoV-2 (motsoako oa RdRp le N gene) ke 87.6%, boleng ba mofuta oa ORF1a/b Ct ea tlhahlobo ea Sansure Biotech 2019-nCoV. e bonts'a Ct 值(<20 Ct) e tlase liperesente tse 50.3%,高Ct 值(36–40 Ct) 的 peresente ke 3.2%. Как показано на рисунке 1, анализ Abbott SARS-CoV-2 (сочетающий гены RdRp и N) имел самое низкое процентное значение Ct (< 20 Ct% , 8 , 8 гена ORF1a/b в исследовании Sansure Biotech 2019- Анализ nCoV показал низкий Ct. Joalo ka ha ho bonts'itsoe ho Setšoantšo sa 1, tlhahlobo ea Abbott SARS-CoV-2 (e kopanyang mefuta ea RdRp le N) e ne e na le liperesente tse tlase tsa Ct boleng (<20 Ct) ho 87.6%, ha boleng ba Ct ba mofuta oa ORF1a/b ho Sansure. Boithuto ba Biotech 2019 - Tlhahlobo ea nCoV e bonts'itse Ct e tlase. Процент значений (< 20 Ct) составил 50,3%, а процент высоких значений Ct (36–40 Ct) составил 3,2%. Peresente ea boleng (<20 Ct) e ne e le 50.3%, 'me peresente ea boleng bo phahameng ba Ct (36-40 Ct) e ne e le 3.2%.Teko ea Abbott SARS-CoV-2 B e tlaleha boleng ba Ct bo kaholimo ho 30. Ka lehlakoreng le leng, ho BGI SARS-CoV-2 mofuta oa ORF1a / b o ne o e-na le boleng bo phahameng ba Ct (> 36 Ct) peresente e ne e le 4% (setšoantšo sa 1). Ka lehlakoreng le leng, ho BGI SARS-CoV-2 mofuta oa ORF1a / b o ne o e-na le boleng bo phahameng ba Ct (> 36 Ct) peresente e ne e le 4% (setšoantšo sa 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). Ka lehlakoreng le leng, tlhahlobong ea mofuta oa BGI SARS-CoV-2 ORF1a/b e ne e e-na le boleng bo phahameng ba Ct (> 36 Ct), peresente ea eona e neng e le 4% (setšoantšo sa 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）. Ka lehlakoreng le leng, tlhahlobong ea BGI SARS-CoV-2, peresente ea mofuta oa ORF1a/b e nang le boleng bo phahameng ba Ct (>36 Ct) ke 4% (Setšoantšo sa 1). С другой стороны, в анализе BGI SARS-CoV-2 процент генов ORF1a/b с высокими значениями Ct (>36 Ct) составил 4% (рис. 1). Ka lehlakoreng le leng, tlhahlobisong ea BGI SARS-CoV-2, peresente ea liphatsa tsa lefutso tsa ORF1a/b tse nang le boleng bo phahameng ba Ct (> 36 Ct) e ne e le 4% (setšoantšo sa 1).
Thutong ena, re nkile mehlala ea 164 ea nasopharyngeal. Bakeng sa mefuta eohle ea liteko, ho itšehla thajana le ho holisa RNA ho ile ha etsoa ho sebelisoa mekhoa le lisebelisoa tse khothaletsoang ke bahlahisi ba amehang.
Boithuto bona bo bonts'itse hore tlhahlobo ea Abbott bakeng sa SARS-CoV-2 e na le ts'ebetso e ts'oanang ea ho lemoha joalo ka CRS, e nang le 100% e ntle, e mpe le ka kakaretso concordance. Tumellano ea Cohen ea kappa ke 1.00, e bontšang tumellano e feletseng le CRS. Patlisiso e ts'oanang e entsoeng ke Univesithi ea Washington e US e fumane hore kutloelo-bohloko le boits'oaro bo felletseng ba tlhahlobo ea Abbott bakeng sa SARS-CoV-2 e ne e le 93% le 100%, ka ho latellana, ha e bapisoa le tlhahlobo e khethiloeng ke laboratoring (LDA) ea CDC. . 11. Sistimi ea ho lemoha ea Abbott SARS-CoV-2 e ipapisitse le tlhahlobo e kopaneng ea nako e le 'ngoe ea liphatsa tsa lefutso tsa N le RdRp, kaha liphatsa tsa lefutso ka bobeli li na le kutloelo-bohloko, ho fokotsa li-negative tsa bohata12. Boithuto bo entsoeng Vienna, Austria le bona bo bonts'itse hore lipampiri tse kholo tsa lisampole tse ntšoang le ho lemoha li-volume tse ngata li fokolitse litlamorao tsa phokotso le ho eketsa ts'ebetso ea ho lemoha13. Kahoo, papali e phethahetseng ea Abbott bakeng sa tlhahlobo ea SARS-CoV-2 e ka amahanngoa le sisteme ea ho lemoha sethala eo ka nako e ts'oanang e fumanang liphatsa tsa lefutso tse kopaneng, e ntša lisampole tse ngata (0.5 ml), mme e sebelisa bongata bo bongata (40 µl).
Liphetho tsa rona li boetse li bontšitse hore ts'ebetso ea ho lemoha ha tlhahlobo ea liphatsa tsa lefutso tsa Daan e ne e batla e tšoana le ea CRS. Sena se lumellana le thuto14 e entsoeng Univesithing ea Anhui e Huainan, Chaena, le tseko ea moetsi ea tumellano e ntle ea 100%. Leha ho na le litlaleho tsa liphetho tse sa fetoheng, sampole e le 'ngoe e ne e le mpe ka mor'a ho etsa tlhahlobo e tšoanang, empa e ne e le ntle litlhahlobong tsa Abbott SARS-CoV-2 le Sansure Biotech nCoV-2019. Sena se fana ka maikutlo a hore ho ka 'na ha e-ba le phapang pakeng tsa liphetho tsa mefuta e fapaneng ea liteko. Leha ho le joalo, thutong e entsoeng China15, sephetho sa tlhahlobo ea Daan Gene se ne se fapane haholo (p <0.05) ha se bapisoa le tlhahlobo ea bona ea litšupiso e hlalositsoeng lab. Leha ho le joalo, thutong e entsoeng China15, sephetho sa tlhahlobo ea Daan Gene se ne se fapane haholo (p <0.05) ha se bapisoa le tlhahlobo ea bona ea litšupiso e hlalositsoeng lab. Тем не менее, в исследовании, проведенном в Китае15, результат анализа Daan Gene значительно отличался (p <0,05) от их лаборанатор. Leha ho le joalo, phuputsong e entsoeng China15, sephetho sa tlhahlobo ea Daan Gene se ne se fapane haholo (p <0.05) ho tsoa tlhahlobong ea bona ea litšupiso tsa laboratori.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. Leha ho le joalo, phuputsong e entsoeng Chaena15, liphello tsa tlhahlobo ea liphatsa tsa lefutso tsa Daan li ne li fapane haholo (p <0.05) ha li bapisoa le tlhahlobo ea laboratori ea litšupiso.Phapang ena e kanna ea bakoa ke kutloelo-bohloko ea tlhahlobo ea litšupiso ho bona SARS-CoV-2, mme lithuto tse ling li ka ba bohlokoa ho fumana sesosa.
Ntle le moo, boithuto ba rona bo hlahlobile ts'ebetso ea papiso ea SARS-CoV-2 BGI assay le CRS, e bonts'a tumellano e ntle ea liperesente tse ntle (PPA = 97.9%), tumellano ea liperesente tse mpe (NPA = 100%), le tumellano ea liperesente ka kakaretso ka bong. OPA). ). = 98.8%). Litekanyetso tsa Cohen's Kappa li bontšitse tumellano e ntle (K = 0.975). Lithuto tsa Netherlands16 le China15 li bonts'itse liphetho tse ts'oanang. Teko ea SARS-CoV-2 BGI ke tlhahlobo ea mofuta o le mong (ORF1a/b) e sebelisang 10 µl amplification/detection eluate. Leha ho na le tumellano e ntle ea lipalo-palo le liphetho tsa rona tsa litšupiso, tlhahlobo e ile ea hloloheloa lisampole tse peli tse ntle (1.22%) tsa sampole eohle. Sena se ka ba le litlamorao tse kholo tsa kliniki bakeng sa matla a phetiso ho mokuli le maemong a sechaba.
Tlhahlobo e 'ngoe ea papiso e kenyellelitsoeng thutong ena e ne e le tlhahlobo ea Sansure Biotech nCoV-2019 rRT-PCR (RUO); kakaretso ea papali e ne e le 96.3%. Matla a tumellano a ne a boetse a khethoa ke boleng ba Cohen's Kappa, e neng e le 0.925, e bontšang tumellano e feletseng le CRS. Hape, liphetho tsa rona li tšoana le liphuputso tse entsoeng Univesithing ea Central South e Changsha, Chaena, le Lefapheng la Clinical Laboratory la Liuzhou People's Hospital, Liuzhou City, China17. Leha ho ile ha tlalehoa tumellano e ntle ea lipalo-palo e kaholimo, tlhahlobo ea chi-square (teko ea MacNemar) e bonts'itse hore sephetho sa tlhahlobo ea Sansure Biotech se bile le phapang e kholo ea lipalo ha e bapisoa le CRS (p <0.005). Leha ho ile ha tlalehoa tumellano e ntle ea lipalo-palo e kaholimo, tlhahlobo ea chi-square (teko ea MacNemar) e bonts'itse hore sephetho sa tlhahlobo ea Sansure Biotech se bile le phapang e kholo ea lipalo ha e bapisoa le CRS (p <0.005). Несмотря на то, что было зафиксировано указанное выше хорошее статистическое соответствие, критерий хи-квадрат (критерий Макнезер, критерий хи-квадрат) Sansure Biotech имеет статистически значимое различие по сравнению с CRS (p <0,005). Le hoja tumellano e ntle ea lipalo-palo e ka holimo e tlalehiloe, tlhahlobo ea chi-square (teko ea McNemar) e bontšitse hore sephetho sa tlhahlobo ea Sansure Biotech se na le phapang e kholo ea lipalo ha e bapisoa le 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 检测的结果会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 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 зназим между анализом Sansure Biotech le CRS. Ho sa tsotellehe tumellano e ntle ea lipalo-palo e boletsoeng ka holimo, tlhahlobo ea chi-square (teko ea McNemar) e bontšitse phapang e kholo ea lipalo (p <0.005) pakeng tsa tlhahlobo ea Sansure Biotech le CRS.Likarolo tse tšeletseng (3.66%) li fumanoe e le li-negative tsa bohata ha li bapisoa le CRS (Tlaleho ea Tlatsetso ea 1); sena se bohlokoa haholo, haholo-holo ho latela matla a phetiso ea vaerase. Lintlha tse ka holimo li boetse li tšehetsa sekhahla sena sa ho lemoha se tlaase15.
Thutong ena, litekanyetso tsa Ct li ile tsa khethoa bakeng sa tlhahlobo ka 'ngoe le sethala se fapaneng, ka boleng bo tlase ba Ct bo tlalehiloeng tekong ea Abbott SARS-CoV-2. Sephetho sena se kanna sa amana le sistimi e kopaneng ea tlhahlobo ea liphatsa tsa lefutso ea Abbott ka nako e le 'ngoe bakeng sa ho lemoha SARS-CoV-2. Ka hona, ho ea ka Setšoantšo sa 1, 87.6% ea liphetho tsa Abbott SARS-CoV-2 li ne li e-na le litekanyetso tsa Ct tse ka tlaase ho 20. Ke palo e fokolang feela ea liphetho tsa sampuli (12.4%) e neng e le 20-30. Litekanyetso tsa Ct tse ka holimo ho 30 ha lia tlalehoa. Ntle le ts'ebeliso ea Abbott ea mofuta oa tlhahlobo ea liphatsa tsa lefutso tsa SARS-CoV-2, sephetho sena se kanna sa amana le moeli o tlase oa tlhahlobo (32.5 RNA copies/mL) 18, e tlase makhetlo a mararo ho feta moeli o tlase oa k'hamphani oa likopi tse 100 tsa RNA. /mL. ml)19.
Phuputso ena e na le mefokolo e itseng: ea pele, ha re na mekhoa e tloaelehileng/ea litšupiso [joalo ka bongata ba vaerase kapa liteko tse ling tsa laboratori (LDA)] ka lebaka la khaello ea lisebelisoa. Ea bobeli, mehlala eohle e sebelisitsoeng thutong ena e ne e le li-nasopharyngeal swabs, athe liphello li ne li sa sebetse mefuteng e meng ea mohlala, 'me ea boraro, boholo ba mohlala oa rona bo ne bo le bonyenyane.
Boithuto bona bo bapisitse ts'ebetso ea liteko tse 'ne tsa rRT-PCR bakeng sa SARS-CoV-2 ho sebelisa lisampole tsa nasopharyngeal. Litlhahlobo tsohle tsa ho lemoha li bile le ts'ebetso e batlang e bapisoa, ntle le tlhahlobo ea Sansure Biotech. Ntle le moo, sekhahla se tlase sa positivity se khethiloe tlhahlobong ea Sansure Biotech ha e bapisoa le CRS (p <0.05). Ntle le moo, sekhahla se tlase sa positivity se khethiloe tlhahlobong ea Sansure Biotech ha e bapisoa le CRS (p <0.05). Кроме того, в тесте Sansure Biotech был выявлен низкий процент положительных результатов по сравнению с CRS (p <0,05). Ho feta moo, tlhahlobo ea Sansure Biotech e bontšitse karolo e tlase ea liphetho tse ntle ha li bapisoa le 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). Ntle le moo, tlhahlobo ea Sansure Biotech e bile le sekhahla se tlase sa positivity ha se bapisoa le CRS (p <0.05).Tlhahlobo ea Sansure Biotech nCoV-2019 (RUO) ea PPA, NPA le tumellano ka kakaretso e fetile 93.5% ka matla a Cohen Kappa a boleng ba tumellano ea 0.925. Qetellong, Sansure Biotech Assay (RUO) e hloka netefatso e eketsehileng hore e sebelisoe Ethiopia, 'me lipatlisiso tse ling li lokela ho nahanoa bakeng sa ho hlahloba likopo tse tsoang ho bahlahisi ka bomong.
Moralo oa thuto ea papiso o entsoe litsing tsa bophelo tse 'ne Addis Ababa, Sepetlele sa Eka Kotebe, Setsi sa Kalafo ea Kereke ea Millennium, Sepetlele sa Zewooditu Memorial, le Sepetlele sa Setsebi sa Lefuba sa St. Peter's Tuberculosis. Lintlha li ile tsa bokelloa pakeng tsa la 1 Tšitoe le la 31, 2020. Litsi tsa bongaka tsa phuputso ena li khethiloe ka boomo ho latela palo ea tsona e phahameng ea linyeoe le boteng ba litsi tse kholo tsa kalafo toropong. Ka mokhoa o ts'oanang, lisebelisoa, ho kenyelletsa le lisebelisoa tsa PCR tsa ABI 7500 le Abbott m2000 tsa nako ea nnete, li khethiloe ho latela likhothaletso tsa baetsi ba reagent ea NAAT, 'me ho ile ha khethoa lisebelisoa tse' nè tsa PCR bakeng sa thuto ena, kaha lilaboratori tse ngata tsa Ethiopia li ne li sebelisa bonyane bonyane. bane ba bona. Teko ea Gene, tlhahlobo ea Abbott SARS-CoV-2, tlhahlobo ea Sansure Biotech, le tlhahlobo ea SARS-CoV-2 BGI e entsoeng nakong ea thuto).
Teko ea SARS-CoV-2 e entsoe ho tloha ka la 1 ho isa la 30 Tšitoe 2020 ho sebelisoa 3 ml ea Viral Transport Medium (VTM) (Miraclean Technology, Shenzhen, China) ho tsoa ho batho ba ntseng ba fuputsoa bakeng sa COVID-19 ba bitsitsoeng EPHI. Mehlala ea Nasopharyngeal e ile ea bokelloa ke babokelli ba koetlisitsoeng 'me ba romeloa ho EPHI ka lipakete tse tharo. Pele ho khethollo ea nucleic acid, sampole ka 'ngoe e fuoa nomoro ea boitsebahatso e ikhethang. Ho ntšoa ho etsoa ho tsoa ho sampole ka 'ngoe hang ha ho fihla ho sebelisoa mekhoa ea matsoho le ea othomathiki. Kahoo, bakeng sa ho ntšoa ka mokhoa o itekanetseng oa Abbott m2000, 1.3 ml (ho kenyeletsoa 0.8 ml ea molumo o shoeleng le 0.5 ml ea molumo oa inlet) oa sampuli e ile ea ntšoa ho tloha sampuling ka 'ngoe 'me ea fetisoa ka Abbott DNA Sample Preparation System (Abbott Molecular Inc. des Plaines, IL, USA). ) Sehlopha sa 96 [lisampole tse 92, li-control tse peli tsa ho lemoha le tse peli tse sa sebelisetsoeng li-template (NTC)] li kenyelelitsoe ts'ebetsong ea kakaretso (ho khutlisa le ho lemoha) ea mekhahlelo e 'meli ea SARS-CoV-2 (EUA) ka nako ea sebele. meepo. Ka mokhoa o ts'oanang, bakeng sa ho ntša ka letsoho, sebelisa lisampole tse tšoanang (bakeng sa ho ntša le ho sibolla ka mokhoa o itekanetseng). Kahoo, nakong eohle ea ts'ebetso, lisampole tsa 140 µl li ile tsa qotsoa le ho ntšoa ho sebelisoa QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Germany) ka lihlopha tsa 24 (ho kenyeletsoa lisampole tse 20, li-control tse peli tsa liteko le li-NTC tse peli) ho feta mekhahlelo e robong. Li-eluate tse nkiloeng ka letsoho li ile tsa holisoa 'me tsa bonoa ho sebelisoa sebaesekele se futhumatsang sa ABI 7500 se sebelisang tlhahlobo ea SARS-CoV-2 BGI, tlhahlobo ea Daan Gene, le tlhahlobo ea Sansure Biotech.
Ho itšehla thajana le tlhoekiso ea SARS-CoV-2 viral RNA e latela molao-motheo oa sefaha sa makenete o sebelisang li-reagents tsa ho lokisa sampole tsa Abbott DNA. Ho sa sebetse ha disampole le solubilization ea likaroloana tsa kokoana-hloko ho etsoa ho sebelisoa sesepa se nang le guanidine isothiocyanate ho senya protheine le ho koala RNase. RNA e ntan'o aroloa ho protheine ka ho arohana ha mekhahlelo e tiileng ho sebelisa silica, ke hore letsoai la guanidinium le pH ea alkaline ea lysis buffer e khothalletsa ho tlama ha nucleic acid ho silika (SiO2). Mohato oa ho hlatsoa o tlosa liprotheine tse setseng le litšila ho hlahisa tharollo e hlakileng. Transparent RNA e arohane le li-microparticles tse thehiloeng ho silika ho sebelisa matla a khoheli a sesebelisoa20,21. Ka lehlakoreng le leng, ho itšehla thajana le ho hloekisoa ha RNA ho etsoa ka mokhoa oa ho pota-pota ho sebelisa centrifugation ho e-na le ho ema ha makenete le ho arohana ha li-microparticles ho tloha ho tse ling.
Teko ea Tlhahlobo ea Abbott ea Nako ea 'Nete ea SARS-CoV-2 (Abbott Molecular, Inc.) e entsoe ho latela litaelo tsa moetsi, tse amohetseng EUA19,22 ho tsoa ho WHO le FDA. Protocol ena, ho se sebetse ha sampole pele ho ntšoa ho ne ho etsoa ka bateng ea metsi ho 56 ° C bakeng sa 30 min. Kamora ho se sebetse ha kokoana-hloko, ho ntšoa ha nucleic acid ho ile ha etsoa sesebelisoa sa Abbott m2000 SP ho tloha ho 0.5 ml VTM ho sebelisa mokhoa oa ho lokisetsa sampole oa Abbott m2000 DNA. ho latela moetsi. Ho phahamisa le ho lemoha ho ile ha etsoa ka sesebelisoa sa Abbott m2000 RT-PCR, 'me ho fumanoa ha bobeli ho ile ha etsoa bakeng sa liphatsa tsa lefutso tsa RdRp le N. ROX) le VIC P (dae e nang le thepa) bakeng sa ho lebisa tlhokomelo le ho lemoha taolo ea ka hare, ho lumella ho lemoha ka nako e le 'ngoe lihlahisoa tse peli tsa amplification 19.
Mokhoa oa ho lemoha ka matlafatso ea kit ena o ipapisitse le theknoloji ea RT-PCR ea mohato o le mong. Liphatsa tsa lefutso tsa ORF1a/b le N li khethiloe e le libaka tse sirelelitsoeng ke Daan Gene Technology ho bona katoloso ea sebaka seo ho shebiloeng ho sona. Li-primers tse khethehileng le li-fluorescent probes (li-N gene probes tse ngotsoeng ka FAM, ORF1a/b probes tse ngotsoeng ka VIC) li etselitsoe ho bona SARS-CoV-2 RNA ka lisampole. Metsoako ea ho qetela e hlakileng le e hloahloa e lokisitsoe ka ho kenyelletsa 5 µl ea eluent ho 20 µl ea master mix ho molumo oa ho qetela oa 25 µl. Ho phahamisa le ho lemoha ho entsoe ka nako e le 'ngoe ho sesebelisoa sa PCR sa nako ea sebele sa ABI 750024.
Liphatsa tsa lefutso tsa ORF1a/b le N li ile tsa bonoa ho sebelisoa Sansure Biotech nCoV-2019 Nucleic Acid Diagnostic Kit (ho lemoha PCR ea fluorescent). Lokisetsa li-probes tse khethehileng bakeng sa mofuta o mong le o mong oa sepheo ka ho khetha mocha oa FAM bakeng sa sebaka sa ORF1a / b le mocha oa ROX oa N gene. Sehlopheng sena sa tlhahlobo, li-eluent le master mix reagent li eketsoa ka tsela e latelang: lokisa 30 µl master mix reagent le 20 µl eluted sampole bakeng sa ho lemoha/ho holisa. PCR ABI 750025 ea nako ea sebele e ne e sebelisetsoa ho hōlisa/ho lemoha.
Teko ea SARS-CoV-2 BGI ke sesebelisoa sa fluorescent sa nako ea nnete sa rRT-PCR bakeng sa tlhahlobo ea COVID-19. Sebaka seo ho shebiloeng ho sona se sebakeng sa ORF1a/b sa genome ea SARS-CoV-2, e leng mokhoa o le mong oa ho lemoha lefutso. Ho feta moo, mofuta oa β-actin oa motho oa ho boloka ntlo ke mofuta oa sepheo o laoloang ka hare. Motsoako o moholo o lokisoa ka ho kopanya 20 µl ea master mix reagent le 10 µl ea sampole ea RNA e ntšitsoeng ka poleiting ea seliba26. Sesebelisoa sa PCR sa ABI 7500 fluorescent quantitative real-time se ile sa sebelisoa bakeng sa ho holisa le ho lemoha. Li-nucleic acid amplification tsohle, PCR e tsamaisa maemo bakeng sa tlhahlobo e 'ngoe le e' ngoe, 'me tlhaloso ea liphetho e entsoe ho latela litaelo tsa moetsi (Letlapa la 3).
Tlhahlobong ena ea papiso, ha rea ​​ka ra sebelisa mokhoa o tloaelehileng oa litšupiso ho fumana tumellano ea liperesente (e ntle, e mpe, le ka kakaretso) le lipapiso tse ling tsa litlhahlobo tse 'ne. Papiso e 'ngoe le e 'ngoe ea teko e entsoe le CRS, thutong ena CRS e ne e behiloe ka molao oa "ntho leha e le efe e ntle" 'me sephetho se ile sa khethoa, eseng ka teko e le' ngoe, re sebelisitse bonyane liphetho tse peli tse tšoanang. Ntle le moo, molemong oa phetiso ea COVID-19, liphetho tse fosahetseng li kotsi ho feta liphetho tse ntle tse fosahetseng. Ka hona, ho re "positive" ka nepo kamoo ho ka khonehang ho tsoa sephethong sa CRS, bonyane liteko tse peli li tlameha ho ba tse ntle, ho bolelang hore bonyane sephetho se le seng se nepahetseng se kanna sa tsoa tekong ea EUA. Kahoo, ho liphetho tse 'ne tsa liteko, liphetho tse peli kapa ho feta tse fanang ka sephetho se tšoanang li nkuoa li le ntle kapa li le mpe18,27.
Lintlha li ile tsa bokelloa ho sebelisoa liforomo tse hlophisitsoeng tsa ho ntša lintlha, ho kenngoa ha data le ho hlahloba ho ile ha etsoa ho sebelisoa Excel statistical software le SPSS version 23.0 bakeng sa lipalo-palo tse hlalosang. Tumellano e ntle, e mpe, le ea kakaretso ea liperesente e ile ea hlahlojoa, 'me ho ile ha sebelisoa lintlha tsa Kappa ho fumana tekanyo ea tumellano ea mokhoa o mong le o mong le CRS. Litekanyetso tsa Kappa li hlalosoa ka tsela e latelang: 0.01 ho 0.20 bakeng sa tumellano e bonolo, 0.21 ho 0.40 bakeng sa tumellano e akaretsang, 0.41-0.60 bakeng sa tumellano e itekanetseng, 0.61-0.80 bakeng sa tumellano e kholo le 0.81-0.99 bakeng sa tumellano e feletseng28.
Ho ile ha fumanoa tumello ea boitšoaro ho tsoa Univesithing ea Addis Ababa 'me liprothokholo tsohle tsa liteko tsa phuputso ena li ile tsa amoheloa ke Boto ea Tlhahlobo ea Boitšoaro ba Saense ea Setsi sa Bophelo bo Botle sa Ethiopia. Nomoro ea litšupiso bakeng sa License ea EPHI Ethics ke EPHI/IRB-279-2020. Mekhoa eohle e sebelisitsoe ho latela likhothaletso le lipehelo tsa Tataiso e Pharalletseng ea Naha ea Ethiopia bakeng sa Phekolo ea COVID-19. Ho phaella moo, tumello e ngotsoeng e nang le tsebo e ile ea fumanoa ho barupeluoa bohle ba thuto pele ba kenya letsoho thutong.
Lintlha tsohle tse fumanoeng kapa tse hlahlobiloeng thutong ena li kenyelelitsoe sehloohong sena se hatisitsoeng. Lintlha tse tšehetsang liphetho tsa phuputso ena li fumaneha ho tsoa ho sengoli se amehang ka kopo e utloahalang.
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