Ukusebenza kwemizamo emine yokukhulisa ye-nucleic acid ukukhomba i-SARS-Cov-2 e-Ethiopia

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Kusukela ngesifo sika-2019 CORONAVIRUS isifo se-coronavirus (COVID-19), izivivinyo eziningi zokugcina ze-nucleic acid amplication (Naats) zithuthukiswe emhlabeni wonke futhi zibe yimicimbi ejwayelekile. Yize izivivinyo eziningana zithuthukiswa ngokushesha futhi zasetshenziswa ekuhlolweni kokuxilonga kwelebhu, ukusebenza kwalezi zivivinyo akuzange kuhlolwe ezinhlobonhlobo zezilungiselelo. Ngakho-ke, lolu cwaningo luhlose ukuhlola ukusebenza kwe-abbott SARS-Cov-2, i-Daan Gene, BGI, kanye ne-sansare biotech Lolu cwaningo lwenziwe kwi-Ethiopian Public Health Institute (EPHI) kusuka ku-1 kuye ku-30 ​​kuDisemba 2020. Offidene 164, ama-59.1% ayenempilo futhi u-40.9% ayengalungile ku-CRS. I-Siotech Biotech Positivity yayiphansi kakhulu ngokuqhathaniswa ne-CRS (P <0.05). I-Siotech Biotech Positivity yayiphansi kakhulu ngokuqhathaniswa ne-CRS (P <0.05). I-Sangulach Biotech xыожтачжтн нине по Сsgentо Сравнениир 0 0 05). Imiphumela emihle ka-Bensach ye-Biotech yayiphansi kakhulu ngokuqhathaniswa ne-CRS (P <0.05).与 Crs 相比, sansare biotech 的阳性率显着较低 (p <0.05).与 Crs 相比, sansare biotech 的阳性率显着较低 (p <0.05). У <0 0 0,05). I-Sature Biotech yayinemiphumela emihle emincane ngokuqhathaniswa ne-CRS (P <0.05).Isivumelwano esiphelele salawo mahlaziya amane sasingu-96.3-100% uma siqhathaniswa ne-CRS. Ngaphezu kwesilinganiso esiphansi se-positicivity se-biotech ye-biotech, ukusebenza kwalezi zindlela ezine kwacishe kuqhathaniswa. Kanjalo, i-biotech ye-sansare [ucwaningo kuphela (Ruo)] Assay idinga ukuqinisekiswa okwengeziwe ukuze kusetshenziswe e-Ethiopia. Ekugcineni, ucwaningo olwengeziwe kufanele lubhekwe ukuze luhlole Izici Zokufakazelwa Komkhiqizi Ofanele.
Ukuhlolwa kwelebhu kuyingxenye ye-World Health Organisation (WHO) Isu Lamasu Lesifo seCoronavirus 2019 (Covid-19) Ukulungela nokuphendula (i-SPRP). Ngubani oweluleka ukuthi amazwe adinga ukwakha umthamo welebhu wokuthuthukisa ukulungela, ukuphathwa kwamacala afanele, ukuphaphama kanye nokuphendula ngokushesha ezinseleleni zezempilo zomphakathi. Lokhu kuphakamisa ukuthi indima yelebhu iyisihluthulelo sokubonisa lesi sifo kanye nobhuka ama-epidemiology wabameli abathathelwanayo abaningi futhi balawule ukusakazeka kwabo.
Ukuxilongwa kwe-Covid-19 kudinga imininingwane yezobusha kanye nezokwelapha, izimpawu zomuntu / izimpawu, kanye nedatha ye-radiographic nelebhularabula. Kusukela kwabikwa e-Covil-19 Ukuqubuka eWuhan, eChina, izivivinyo eziningi zokumelana ne-nucleic acid ze-nucleic acid (uNahaats) athuthukisiwe emhlabeni jikelele. I-Polymerase Reverse Reverse Presment Reaction (RRT-PCR) isetshenzisiwe njengendlela ejwayelekile neyejwayelekile yokuxilongwa kwelebhu okuthe xaxa kwe-acute respiratory syndrome 2 (SARS-Cov-2) 3 Ukutheleleka. Ukutholwa kwamangqamuzana kwe-SARS-Cov-2 kuvame ukususelwa ku-n (genein genein protein), ne-RNA (RNA-Uncike e-Pelmerase Gene) Uhlobo lwe-ORF1A / B (Uhlaka Lokufunda Oluvulekile 1A / B). IGene) esifundeni esikhonjwe eViral Genome. Babhekwa njengezifunda eziphambili ezilondoloziwe ezitholakala ku-Viral GetES for virus ukuqashelwa4. Phakathi kwalezi zigaba zofuzo, i-RDRP ne-E NEZENSE LUNGANI UKUTHOLAKALA KWEMPAHLA YOKUQALA, kuyilapho i-N Gene inozwela oluphansi lokuhlaziya.
Ukusebenza kwe-PCR ASTS kungahluka ngokuya ngezinto ezahlukahlukene ezifana ne: Ukukhishwa kwama-reagents, ama-amplification / ukutholwa kwemvume, indlela yokukhipha, ikhwalithi yomshini we-PCR nezinye izinsimbi. Kusukela ngo-Ephreli 2020, amadivaysi angaphezu kwama-48 anqamula avela emazweni ayisishiyagalolunye athole ukugunyazwa okusetshenziswa kwezimo eziphuthumayo (eua) ngokuxilongwa kwe-Covil-196. E-Ethiopia, amapulatifomu e-PCR angaphezu kwe-14 asetshenziselwa ukutholwa kwe-PCR-HOL-2 ezikhungweni zezempilo zomphakathi ezingama-26, kufaka phakathi i-ABI 7500, i-ABBHE 48000 ne-STUDIO7. Ngaphezu kwalokho, amakhithi ahlukahlukene wokuhlola we-PCR ayatholakala, anjenge-Daan Gene Test, Abbott Sars-Cov-2 Test, ukuhlolwa kwe-biotech, kanye ne-SARS-Cov-2-2 BGE Test. Although rRT-PCR is highly sensitive, some patients with COVID-19 report false negative results due to insufficient copies of viral ribonucleic acid (RNA) in samples due to improper collection, transport, storage and handling, and laboratory testing. izimo nezenzo zabasebenzi8. Ngaphezu kwalokho, isampula noma i-mishandling yesampula, i-mishandling, umjikelezo womjikelezo (i-CT), futhi unqamule ukuwela namanye ama-pathogenic nucleic acid noma ama-SARS / asalayo we-SARS-2 Ngakho-ke, kuyacaca ukuthi ukuhlolwa kwe-PCR kungakhomba izinto ezithwala izinto zofuzo, ngoba zingenakukwazi ngisho nokuhlukanisa phakathi kwezizukulwane ezisebenzayo zegciwane, ngakho-ke izivivinyo zingakhomba kuphela abathwali hhayi iziguli. Ngakho-ke, kubalulekile ukuhlola ukusebenza kokuxilonga kusetshenziswa izindlela ezijwayelekile esimweni sethu. Yize ama-naat amaningi ama-reagents atholakala kwi-Ethiopian Public Health Institute (EPHI) nakuwo wonke amazwe, akukho ukuhlolwa kokuqhathanisa nokuphumelela kwawo kungakabikwa. Ngakho-ke, lolu cwaningo luhlose ukuhlola ukusebenza kokuqhathanisa kwamakhithi atholakala kwezentengiso wokutholwa kwe-SARS-CoV-2 nge-RRT-PCR ngokusebenzisa ama-specimens emitholampilo.
Bangu-164 abahlanganyeli ababambiqhaza abasolwa ngokusola abangu-19 bafakiwe kulolu cwaningo. Iningi lamasampula lalivela ezikhungweni zokwelashwa (118/164 = 72%), ngenkathi ababambiqhaza abangama-46 (28%) abasele bebezokwelashwa. Phakathi kwababambiqhaza abangazange baphathwe esikhungweni, esingu-15 (9.1%) basolwa ngamacala asolwa ngokomtholampilo kanye nama-31 (18.9%) abaxhumana nabo abaqinisekisiwe. Ababambiqhaza abangamashumi ayisishiyagalolunye nantathu (56.7%) babengowesilisa, kanye ne-SD (± SD) yobudala babahlanganyeli kwakuyiminyaka engama-3110 (± 11.82).
Kulolu cwaningo, amanani amahle nezimbangele ezivivinyo ezine ze-19 ze-19 anqunywa. Ngakho-ke, amanani amahle e-abbott SARS-Cov-2 Assay, Daan Gene 2019-NCOV-Assay, SARS-NCOV-2 BGI Assay, kanye ne-58,5%, 57,5%, 57,5% kanye ne-55.5% ngokulandelana. Izikolo ezi-Creference ezihle kakhulu nezingalungile ze-Composite Crefetha ezijwayelekile (CRS) zazingama-97 (59.1%) no-67 (40.9%), ngokulandelana (itafula 1). Kulolu cwaningo, incazelo ye-CRS yayisuselwa "kunoma yimuphi umthetho omuhle", lapho imiphumela emine yokuhlola, imiphumela emibili noma ngaphezulu yokuhlola enikeze imiphumela efanayo yabhekwa njengeqiniso.
Kulolu cwaningo, sathola isivumelwano esingesihle samaphesenti (i-NPA) ka-100% (95% CI 94.6-100) kubo bonke ukuhlaziya okuqhathaniswa no-CRS. Ukuhlaziywa kwe-biotechnology ye-snsally kwakhombisa i-PPA encane ye-93.8% (95% CI 87.2-97.1) kanye nokuhlaziywa kwe-Daan gene 2019-NCOV kwaba nesivumelwano sonke se-99.4% CI 96.6-99.9). Ngokuphambene, isivumelwano sonke phakathi kwe-SARS-Cov-2 BGI Assay kanye ne-Biotech Biotech 2019-NCOV Assay yayingama-98.8% no-96.3%, ngokulandelana (itafula 2).
I-Cohen's Kappa Coefficient Yesivumelwano phakathi kwe-CRS kanye ne-ABBOTTS SARS-Cov-2 Assay imiphumela yazingaguquki ngokuphelele (k = 1.00). Ngokufanayo, amanani we-kappa kaCoun Gene atholwe nguDaan Gene 2019-NCOV, SARS-Cov-2 BGI, kanye ne-sansare Biotech 2019-Ncov futhi ahambisana ngokuphelele ne-CRS (K ≥ 0.925). Kulokhu kuhlaziywa kokuqhathanisa, ukuhlolwa kwe-Chi-Square (ukuhlolwa kwesikwele
Njengoba kukhonjisiwe ku-Fig.1 Iphesenti lenani eliphansi le-CT (<20 ct) le-Abbott SARS-Cov-2 Assay (i-RDP ehlanganisiwe ne-n gene) yayinenani lamaphesenti angama-87.6 ct) lalingu-50.3% kanye nenani eliphakeme le-CT (36-40 CT) lalingu-3.2%. 1 Iphesenti lenani eliphansi le-CT (<20 ct) le-Abbott SARS-Cov-2 Assay (i-RDP ehlanganisiwe ne-n gene) yayinenani lamaphesenti angama-87.6 ct) lalingu-50.3% kanye nenani eliphakeme le-CT (36-40 CT) lalingu-3.2%.Njengoba kukhonjisiwe ku-Fig.1, зроцент наииимешния CT (<20 ct) Анализа Orf1A / B Аналилиза sansarech 2019-NCOV пока проц чока проц Составлял ama-3,2%. 1, iphesenti lenani eliphansi kakhulu le-CT (<20 ct) lokuhlaziywa kwe-Abbott SARS 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) 值显示低 87.6%, b 基因 ct 值显示低 ct Njengoba kukhonjisiwe kuMdwebo 1, amaphesenti we-CT aphansi kakhulu (<20 ct) wokuhlolwa kwe-Abbott SARS (inhlanganisela ye-RTP ne-NCe 3.2%. К акказ »Abbott SARS-COV-2 (с CT генени А Con генени CT геанени е " Njengoba kukhonjisiwe kuMdwebo 1, i-Abbott SARS-COV-2 Assay (ehlanganisa i-RDP ne-N Gene) yayinenani eliphansi kakhulu le-CT (<20 ct) ngo-87.6%, isifundo se-NCOV sikhombise i-CT ephansi. I-O0,3%, '<20) зы,чхх (( Amaphesenti amanani (<20 ct) ayengama-50.3%, kanye namaphesenti amanani aphezulu we-CT (36-40 CT) kwakungu-3.2%.I-abbott SARS-Cov-2 B Hlola amanani we-CT abhalwe ngenhla 30. Ngakolunye uhlangothi, kwi-BGI SARS-Cov-2 Assay Orf1a / B Uhlobo lwe-CT lube nenani eliphakeme le-CT (> 36 CT) Amaphesenti angu-4% (Fig. 1). Ngakolunye uhlangothi, kwi-BGI SARS-Cov-2 Assay Orf1a / B Uhlobo lwe-CT lube nenani eliphakeme le-CT (> 36 CT) Amaphesenti angu-4% (Fig. 1). Kis доuро4 Стороны, bheka BGI Sars-Cov-2 Ngakolunye uhlangothi, ekuhlaziyeni kwe-BGI Sars-Cov-2 gene Orf1A / B kwakunenani eliphakeme le-CT (> 36 ct), iphesenti lalingu-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). Ngakolunye uhlangothi, ku-BGI Sars-Cov-2 ukutholwa, iphesenti le-Orf1A / B Gene ngenani eliphakeme le-CT (> 36 CT) lingu-4% (Umdwebo 1). Kis доuро4 стоuроны, bheka BGI Sars-Cov-2-2-2 - 2 - 2 Ngakolunye uhlangothi, ekuhlaziyeni kwe-BGI Sars-Cov-2, iphesenti le-Orf1A / B ufuzo ngamanani aphezulu we-CT (> 36 CT) kwakungu-4% (Fig. 1).
Kulolu cwaningo, sathatha amasampula ayi-164 eNasopharyngeal. Kuzo zonke izinhlobo zemizamo, kwenziwa isonto se-RNA kanye nokukhuliswa kwenziwa kusetshenziswa izindlela namakhithi anconywe abakhiqizi abafanele.
Lolu cwaningo lubonise ukuthi ukuhlolwa kuka-Abbott kwe-SARS-Cov-2 kunokusebenza okufanayo kokutholwa njenge-CRS, nge-100% enhle, engemihle, kanye ne-canclackan ephelele. Isivumelwano seKappa sikaCohen singu-1.00, sibonisa isivumelwano esigcwele nge-CRS. Ucwaningo olufanayo ne-University of Washington e-US lwathola ukuthi ukuzwela okuphelele nokucaciswa kokuhlolwa kwe-ABBOTTS kwa-SARS-CoV-2 kwakungu-93% kanye ne-100% ye-CDC (LDA) ye-CDC. 11 Isifundo eVienna, i-Austria ibuye ikhombise ukuthi amavolumu amakhulu asampula kanye namavolumu angu-Elume ancishisiwe anciphise imiphumela yokuhlanza kanye nokwanda kokuthola kabusha13. Ngakho-ke, umdlalo ophelele ka-Abbott we-SARS-CoV-2 Assay angahlotshaniswa nohlelo lokutholwa kweplatifomu okutholakale ngasikhathi sinye, ekhipha inani elikhulu lamasampula (0.5 ml), futhi lisebenzisa inani elikhulu le-eluent (40 μL).
Imiphumela yethu ibuye ikhombise ukuthi ukusebenza kokutholwa kofuzo lwezakhi zofuzo lwe-daan kwakucishe kufane naleyo ye-CRS. Lokhu kuyahambisana nocwaningo14 olwenziwe e-Anhui University eHuinan, eChina kanye nesimangalo somenzi wesivumelwano esihle. Naphezu kwemibiko yemiphumela engaguquki, isampula eyodwa ibingalungile nge-Heath ngemuva kokuthola imali efanayo, kepha yayinethemba e-Abbott SARSS-Cov-2 kanye ne-SATURE BIOTECH NCOV-2019 ASTS. Lokhu kuphakamisa ukuthi kungahle kube nokuhlukahluka kwemiphumela ezinhlotsheni ezahlukahlukene ze-ASAS. Noma kunjalo, ocwaningweni olwenziwe eChina15, umphumela we-Daan gene Assay wawuhluke kakhulu (p <0.05) ngokuqhathaniswa nezembatho zazo ezichazwe nge-lab. Noma kunjalo, ocwaningweni olwenziwe eChina15, umphumela we-Daan gene Assay wawuhluke kakhulu (p <0.05) ngokuqhathaniswa nezembatho zazo ezichazwe nge-lab. Yebo, bheka, Китаедани, К К Ктаеsob, кезулулье15, латарного эталоного Аналинго Анлонгиа. Kodwa-ke, ocwaningweni lwase-China15, umphumela wokuhlaziywa kukaGene we-Daan wawuhluke kakhulu (p <0.05) ekuhlaziyweni kwesethenjwa sabo.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 Yebo, К << I-лэтабоныs. Kodwa-ke, ocwaningweni lwaseChina15, imiphumela yokuhlolwa kofuzo lukaDaya yayihluke kakhulu (p <0.05) ngokuqhathaniswa nokuhlolwa kwe-reference elether.Lokhu kungafani kungahle kube ngenxa yokuzwela kokuhlolwa kwereferensi ukuthola i-SARS-Cov-2, futhi ezinye izifundo zingabalulekile ukuthola imbangela.
Ngaphezu kwalokho, isifundo sethu sahlola ukusebenza kokuqhathanisa kwe-SARS-Cov-2 BGI Assay nge-CRS, kukhombisa isivumelwano esihle samaphesenti (i-PPA = 97.9%), Isivumelwano se-NPA = 100%), kanye nesivumelwano se-Generaji). ). = 98.8%). Amanani weKappa's Kappa akhombisa isivumelwano esihle (k = 0.975). Izifundo eNetherlands16 nase-China15 zikhombise imiphumela engaguquki. Ukuhlolwa kwe-SARS-Cov-2 BGI kuyinto yofuzo eyodwa (ORF1A / B) Ukuhlolwa kokutholwa kusetshenziswa i-10 μl amplification / quecte. Naphezu kwesivumelwano esihle sezibalo nemiphumela yethu yenkomba, ukuhlaziya kwaphuthelwa amasampula amabili amahle (1,22%) wesampula ephelele. Lokhu kungaba nemiphumela emikhulu yemitholampilo yokudlulisela amandla ashukumisayo kuwo womabili amazinga wesiguli kanye nomphakathi.
Okunye ukudingidwa kokuqhathanisa kufakiwe kulolu cwaningo kwakuyi-biotech biotech ncov-2019 RRT-PCR (RUO) Assay; Amaphesenti womdlalo jikelele ayengu-96.3%. Amandla esivumelwano nawo anqunywa yinani le-kappa le-kappa, elalingu-0.925, elibonisa isivumelwano esigcwele ne-CRS. Futhi, imiphumela yethu ifana nezifundo ezenziwa eCentral South University eChangsha, eChina, naseMnyangweni Wezokuhlinzekelwa Kwelebhu Yasemitholampilo yaseLiuzhou People's Hospital, iLiuzhou City, China City, China. Noma kwabhalwa phansi okungenhla kwe-concordance ye-Chistical Concondance, isivivinyo se-Chi-Square (Test Macnemar) sibonisa ukuthi umphumela we-biotech biotech stusy ake waba nomehluko obalulekile ngokwezibalo uma uqhathaniswa ne-CRS (P <0.0000). Noma kwabhalwa phansi okungenhla kwe-concordance ye-Chistical Concondance, isivivinyo se-Chi-Square (Test Macnemar) sibonisa ukuthi umphumela we-biotech biotech stusy ake waba nomehluko obalulekile ngokwezibalo uma uqhathaniswa ne-CRS (P <0.0000). Ezama-Isilinganisweni Sett, новат, (Критууулататзазай Анаtureмара I-0,005). Yize kubhalwe isivumelwano esihle sezibalo, ukuhlolwa kwe-Chi-Square (ukuhlolwa kwesikwele) kukhombisa ukuthi umphumela we-biotech biotech yesistimu ethola umehluko obalulekile ngokuqhathaniswa ne-CRS (P <0.0000).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, sansare biotech 检测的结果与 CRS 相比具有统计学显着差异 (p <0.0000).尽管 记录 了 上述 良好 统计 一致性, 但 检验 ((Macnemar 检验 表明 ,, Sinsare Biotech 检测 结果 与 CRS 相比 具有 显着 ((P <0.0055 ...........))) Ukuze uthole ukwaziswa okwengeziwe, bheka Sх I-Soutитчит Ngaphandle kwesivumelwano esihle sesibalo esishiwo ngenhla, ukuhlolwa kwe-Chi-Square (ukuhlolwa kwe-McNemar) kukhombisa umehluko obalulekile ngokwezibalo (P <0.005) phakathi kwe-biotech biotech ye-motech ye-snsach assay kanye nama-CRS.Amasampula ayisithupha (ama-3.66%) atholwe angabi namanga aqhathaniswa nama-CRS (itafula elengeziwe 1); Lokhu kubaluleke kakhulu, ikakhulukazi okunikezwa amandla okudlulisela kwegciwane. Le mininingwane engenhla ibuye isekela lesi silinganiso esiphansi sokutholwa15.
Kulolu cwaningo, amanani we-CT anqunywe nge-assay ngayinye kanye nesikhulumi esifanele, nenani eliphansi kakhulu le-CT elibikwe ku-ABBOTT SARS-CoV-2 Assay. Lo mphumela ungahle uhlobane nohlelo lokuhlola lofuzo luka-Abbott ngasikhathi sinye sokutholwa kwe-SARS-CoV-2. Ngakho-ke, ngokusho komdwebo 1, 87.6% we-Abbott Sars-Cov-2 imiphumela yayinamanani we-CT ngaphansi kwama-20. Amanani we-CT ngenhla kwama-30 awaqoshwanga. Ngaphezu kokusetshenziswa kuka-Abbott kwefomethi ye-SARS-Cov-2 Panel ml) 19.
Lolu cwaningo lunokulinganiselwa okuthile: Okokuqala, asinazo izindlela ezijwayelekile / zereferensi [njengomthwalo wegciwane noma ezinye izivivinyo zelebhu noma ezinye izivivinyo zelebhu (LDA)] ngenxa yokuntuleka kwezinsizakusebenza. Okwesibili, zonke izinhlobo ezisetshenzisiwe kulolu cwaningo zazingama-nasopharyngeal swabs, kanti imiphumela ingasebenzi kwezinye izinhlobo zezinhlobo, neyesithathu, usayizi wethu wesampula wawumncane.
Lolu cwaningo luqhathanise ukusebenza kwezitholelo ezine ze-RRT-PCR ze-SARS-Cov-2 usebenzisa amasampula we-nasopharyngeal. Zonke izincazelo zokutholwa zazinokusebenza cishe, ngaphandle kwe-biotech ye-biotech ye-assay. Ngaphandle kwalokho, inani eliphansi le-positivity likhonjwe ku-biotech biotech le-stach assay uma kuqhathaniswa ne-CRS (P <0.05). Ngaphandle kwalokho, inani eliphansi le-positivity likhonjwe ku-biotech biotech le-stach assay uma kuqhathaniswa ne-CRS (P <0.05). I-Biotech Biotech äыленняле сравнениии Ngaphezu kwalokho, ukuhlolwa kwe-moitech kwe-moitech kukhombisa amaphesenti aphansi wemiphumela emihle kuqhathaniswa ne-CRS (P <0.05).此外, 与 CRS 相比, sansare biotech 检测的阳性率较低 (p <0.05).此外, 与 CRS 相比, sansare biotech 检测的阳性率较低 (p <0.05). С <0 0 0 0 05). Ngaphezu kwalokho, i-biotech ye-biotech ye-sansare inesilinganiso esisezingeni eliphansi ngokuqhathaniswa ne-CRS (P <0.05).I-Biotech BOITECH NCOV-2019 (Ruo) Ukuhlaziywa kwe-PPA, i-NPA kanye nesivumelwano se-93.5% nge-Cohen Kappa Amandla wenani lesivumelwano le-0.925. Ekugcineni, i-biotech biotech assay (RUO) idinga ukuqinisekiswa okwengeziwe ukuze isetshenziswe e-Ethiopia, futhi ucwaningo olwengeziwe kufanele lubhekwe njengokuhlola izimangalo kubakhiqizi abathile.
Kwenziwe ukwakheka okufundwayo kokuqhathanisa ezikhungweni ezine zezempilo e-Addis Ababa, Isibhedlela sase-Eka Kotebe, Isibhedlela Sokulawulwa Kwezinhlanga Millenitu, kanye nesibhedlela saseSt Peter Tuberculosis. Imininingwane yaqoqwa phakathi kukaDisemba 1 no-31, 2020. Izikhungo zezokwelapha zalolu cwaningo zazikhethwa ngobuqotho ngokuya ngesibalo sazo esiphakeme samacala nokutholakala kwezikhungo ezinkulu zokwelapha edolobheni. Ngokufanayo, izinsimbi, kufaka phakathi i-ABI 7500 kanye ne-abbott m2000 amathuluzi we-real-time PCR, akhethelwa izincomo zabakhiqizi be-naaat realent, kanti amakhithi amane okutholwa kwe-PCR akhethiwe kulolu cwaningo, njengoba iningi lawo malini e-PCRE Ukuhlolwa koGene, ukuhlolwa kwe-Abbott Sars-Cov-2, ukuhlolwa kwe-biotech, kanye ne-SARS-Cov-2 2 BGI kwenziwa ngesikhathi sokufunda).
Ukuhlola i-SARS-Cov-2 kwenziwa kusuka ku-1 kuye ku-30 ​​kuDisemba 2020 kusetshenziswa ama-3 ml we-viral Transport Medium Medium (VTM) (ubuchwepheshe be-Miraclean, iShenzhen, China) kusuka ku-EPHI. Amasampula we-nasopharyngeal aqoqwa ngabathelisi abaqeqeshiwe abaqeqeshiwe futhi athunyelwa e-EPHI ngamaphakethe kathathu. Ngaphambi kokuhlukaniswa kwe-nucleic acid, isampula ngayinye yabelwa inombolo eyingqayizivele kamazisi. I-Extraction yenziwa ekhasini ngalinye ngokushesha lapho kufika izindlela ezisetshenziswayo zokukhishwa kwezandla nezitholile. Ngakho-ke, ngokukhishwa okuzenzakalelayo kwe-abbott m2000, 1.3 ml (kufaka phakathi ivolumu yevolumu engu-0.8 ml kanye ne-0.5 ml yevolumu ye-0.5 ml ) Iqoqo lamasampula angama-96 [92, izilawuli ezimbili zokutholwa nezilawuli ezimbili ezingezona isifanekiso] zifakiwe kwinqubo ephelele (i-NTC)] ifakiwe kwinqubo ephelele (ukubuyisa kanye nokutholwa) kwemijikelezo emibili ye-SARS-CoV-2 (EUA) ngesikhathi sangempela. Imayini. Ngokufanayo, ukukhishwa ngesandla, sebenzisa amasampula afanayo (ngokukhishwa okuzenzakalelayo nokutholwa). Ngakho-ke, kuyo yonke le nqubo, amasampula angu-140 μl awatholakali futhi akhishwe kusetshenziswa i-Qiamp GmbH RNA Mini Kit (qiagen GmbH, Hilden, Germany) kuma-batch angama-24 (kufaka phakathi amasampula angama-20, izilawuli ezimbili ze-Assay) ngaphezulu kwemijikelezo eyisishiyagalolunye. Kutholwe ama-expeed ngesandla ngesandla futhi kutholakele kusetshenziswa i-Abi 7500 cypeer eshisayo esebenzisa i-SARS-Cov-2 BGI Assay, iDaan Gene Assay, kanye neSinsare Biotech Assay.
Ukuhlukaniswa okuzenzakalelayo kanye nokuhlanzwa kwe-SARS-Cov-2 Viral RNA kulandela umgomo wobusizi obunzima usebenzisa i-abbott DNA Sampula Ukulungiswa Kwesampula Ukulungiselela ama-reagents. Ukungasebenzi kwamasampula kanye nokunyakaza kwezinhlayiya zegciwane kwenziwa kusetshenziswa okokuhlanza okuqukethe i-guanidine isothiocyanate ku-detatuin kanye ne-inactivate rnase. I-RNA bese ihlukaniswa namaprotheni ngesigaba sesigaba esiqinile usebenzisa i-silica, okungukuthi usawoti we-guanidium kanye ne-alkaline PH ye-lysis buffer igqugquzele ukubopha ama-nucleic acid ku-silica (Sio2). Isinyathelo se-rinsing sisusa amaprotheni asele kanye nemfucumfucu ukukhiqiza isixazululo esicacile. I-Transparent RNA isodwa kusuka kuma-microparticles asuselwa ku-silica usebenzisa i-magnetic field202. Ngakolunye uhlangothi, ukwahlukaniswa kwezandla kanye nokuhlanzwa kwe-RNA kwenziwa yi-Spin Colummus Way kusetshenziswa i-CentriFugation esebenzisa i-centrifugation esikhundleni sokuma kazibuthe kanye nokwehlukanisa ama-microparticles kusuka ku-exent.
I-abbott real-time sars-cov-2 Ukuhlolwa kokutholwa (i-abbott molecular, Inc.) yenziwe ngokuya ngemiyalo yomenzi, eyathola i-EUA19,22 kusuka kubani ne-FDA. Kulesi protocol, isampula yokungasebenzi ngaphambi kokukhishwa kwenziwa kubhavu wamanzi ku-56 ° C imizuzu engama-30. Ngemuva kwe-Intactivation ye-Virus, ukukhishwa kwe-nucleic acid kwenziwa ku-abbott m2000strest yethuluzi kusuka ku-0.5 m2 vtm usebenzisa uhlelo lwe-abbott M2000 DNA Sampula Ukulungiselela uhlelo. ngokusho komenzi. I-Amplification and Detection yenziwa kusetshenziswa ithuluzi le-ABBOTT M2000 RT-PR-PCR, kanye nokutholwa okumbaxambili kwenziwa nge-RDRP ne-N Genes. I-ROX) kanye ne-VIC P (DEPRIetary D (DEPRIetary D (DEPRIetary D (POSTRIetary D (Ukutholwa kokulawulwa kwangaphakathi, ukuvumela ukutholwa kanyekanye ngasikhathi sinye kwemikhiqizo ye-Amplification 19.
Indlela yokutholwa ye-Amplification yale khithi isuselwa kubuchwepheshe be-RT-STEP BCR eyodwa. I-ORF1A / B ne-N Gen Ama-prebers athile nezingcweti ze-fluorescent (n kungenzeka abhekwe nge-Fam, Orf1a / B Amabheki abhalwe nge-VIC) aklanyelwe ukuthola i-SARS-CoV-2 RNA kumasampula. Ama-eluent a-eluent kanye nama-master ahlanganisiwe alungiswe ngokungeza ama-5 μL e-Eluent kuya ku-20 μL we-Master ukuxubana nevolumu yokugcina engu-25 μl. I-Amplification and Detection yenziwa ngasikhathi sinye kudivayisi ye-ABI 750024 real-time PCR.
I-ORF1A / B ne-N Gen Lungiselela ama-genes athile we-gene gene ngayinye ngokukhetha isiteshi se-FAM ngesifunda se-ORF1A / B kanye nesiteshi se-ROX se-O gene. Kule Kit ye-Assay Kit, i-Eluent kanye ne-Master Mix reagents yengezwa kanjena: Lungiselela i-30 μL Master Master Mix Reagent kanye ne-20 μL enyusiwe isampula yokutholwa / ukukhuliswa. I-PRCH-time PCR ABI 750025 isetshenziselwe ukukhuliswa / ukutholwa.
Isivivinyo se-SARS-Cov-2 BGI yikhithi ye-fluorescent real-time phr Isifunda esiqondiwe sisesifundeni se-ORF1A / B se-SARS-Cov-2 Genome, okuyindlela eyodwa yokutholwa kofuzo. Ngaphezu kwalokho, i-geneving yomuntu β-actin inguhlobo oluqondiswe ngaphakathi. I-Master Mix ilungiselelwe ngokuxuba ama-20 μL we-Master Mix Reagent kanye ne-10 μL yesampula ye-RNA ekhishwe endaweni ye-Plate26. Ithuluzi le-PRCR ye-PRCR yesikhathi sangempela se-ABI 7500 selisetshenziselwa ukukhuliswa kanye nokutholwa. Yonke imephu ye-nucleic acid, izimo zokugijima ze-PCR ye-assay ngayinye, nokutolika kwenziwa ngemiphumela ngokuya ngemiyalo yomkhiqizi womkhiqizi ofanele (Ithebula 3).
Kulokhu kuhlaziywa kokuqhathanisa, asizange sisebenzise indlela ejwayelekile yesethenjwa ukuthola isivumelwano samaphesenti (esimisiwe, esingesihle, futhi sisonke) kanye neminye imingcele yokuqhathanisa yohlaziyi olune. Ukuqhathanisa ngakunye kwe-CRS kwenziwa nge-CRS, kulolu cwaningo ama-CRS abekwe ngumthetho "noma yikuphi okuhle" futhi umphumela wawunqunywa, hhayi ngovivinyo olulodwa lokuhlola. Ngaphezu kwalokho, endabeni yokudluliselwa kwe-Avil-19, imiphumela emibi eyingozi iyingozi kakhulu kunemiphumela emihle yamanga. Ngakho-ke, ukusho ukuthi "okuhle" ngokunemba ngangokunokwenzeka kusuka kumphumela we-CRS, okungenani izivivinyo ezimbili zohlobo kumele zibe zinhle, okusho ukuthi okungenani umphumela owodwa omuhle kungenzeka uqhamuka e-EUA Assay. Ngakho-ke, ngaphandle kwemiphumela emine yokuhlola, imiphumela emibili noma ngaphezulu yokuhlola enikeza umphumela ofanayo kubhekwa njengeqiniso noma i-hif18,27.
Idatha iqoqwe kusetshenziswa amafomu okukhishwa kwedatha ehlelekile, ukungena kwedatha nokuhlaziywa kwenziwa kusetshenziswa isoftware yezibalo ze-Excel Statistical kanye ne-SPSS version 23.0 yezibalo ezichazayo. Kuhlaziywa isivumelwano esibi, esingesihle, futhi sonke isikhathi, kanti nesikolo seKappa sasisetshenziselwa ukuthola izinga lesivumelwano sendlela ngayinye nge-CRS. Amanani we-KAPPA ahunyushwa kanjena: 0.01 kuya ku-0,20 ngesivumelwano esithambile, 0.21 kuya ku-0.40 esivumelwaneni esijwayelekile, 0.41-0.80 esivumelwaneni esikhulu kanye no-0.81-0.99 ngesivumelwano esiphelele28.
Ukuvuselelwa kokuziphatha kwatholakala e-University of Addis Ababa futhi zonke izinqubo zokuhlola zalolu cwaningo zivunywa yi-Ethiopian Public Health Institute Bhorication Bhokisi. Inombolo yesethenjwa yelayisense le-EPHI Ethis yi-EPHI / IRB-279-2020. Zonke izindlela zazisetshenziswa ngokuya ngezincomo nezinhlinzeko zemihlahlandlela kazwelonke kazwelonke yokwelashwa kwe-Covil-19. Ngaphezu kwalokho, imvume ebhaliwe enolwazi yatholwa kubo bonke ababambe iqhaza ocwaningweni ngaphambi kokuzibandakanya ocwaningweni.
Yonke imininingwane etholakele noma ehlaziywe kulolu cwaningo ifakiwe kule ndaba eshicilelwe. Idatha exhasa imiphumela yalolu cwaningo iyatholakala kumlobi ofanele kwisicelo esifanele.
I-World Health Organization. Iziphakamiso zamasu wokuhlola abase-laboratory we-Covid-19: Isiqondisi sesikhashana, Mashi 21, 2020 No. Who / 2012-NCOV_TY_TESTING / 2020.1 (WHO, 2020.1 (WHO, 2020.1 (WHO, 2020).
I-Mouliou, DS, Pantazopoulos, i. & Gourgoulianis, i-COVIK-19 Ukuxilongwa smart emnyangweni wezimo eziphuthumayo: konke-kusebenza. I-Mouliou, DS, Pantazopoulos, i. & Gourgoulianis, i-COVIK-19 Ukuxilongwa smart emnyangweni wezimo eziphuthumayo: konke-kusebenza.I-Muliou, DS, Pantazopoulos, Mina. And Gurgulianis, Ukuxilongwa kwe-Ki ngokuqondile kwe-Covid-19 eMnyangweni Wezimo Eziphuthumayo: konke okusebenzayo.I-Muliou DS, Pantazopoulos I. kanye nokuxilongwa kwe-Gurgilyanis Ki Ki-Ki Intelligent enengqondo ye-Covid-19 eminyangweni yezimo eziphuthumayo: ukuhlanganiswa kokuphela kokuphela kokusebenza. Ukuphefumula uchwepheshe. umuthi. 3, 263-272 (2022).
UMitchell, SL & ST George, K. Ukuvalwa kwe-ID ye-Covid19 manje eua assay. UMitchell, SL & ST George, K. Ukuvalwa kwe-ID ye-Covid19 manje eua assay.UMitchell, SL noSt George, K. Ukuhlaziywa kwe-ID ye-Covid19 manje eua assay.UMitchell SL noSt George K. Ukuvalwa kwe-ID ye-Covid19 manje eua assay. J. Clinical. Igciwane. 128, 104429. Https://doi.org/10.1016/J.JCV.2020.104429 (2020).
I-WHO. Ukutholwa kwelebhu ye-coronavirus isifo sika-2019 (Covid-19) kwisifo somuntu esisolwa ngaso. https://www.who.int/Publications/i/item/10665-331501 (Kufinyelelwe 15 Agasti 20 2020) (WHO, 2020).
UDugama, B. et al. I-Covil-19 Ukuxilongwa: Izifo namathuluzi okuhlola. ACS Nano 14 (4), 3822-3835 (2020).
USyed S. et al. Ukusungulwa kweKholeji Yama-Pathologists aseMpumalanga, Central and Southern Africa - Isikole Sesifunda Sokufundisa kwe-Middle East naseNingizimu Afrika. I-Afrika. J. Lab. umuthi. 9 (1), 1-8 (2020).
Institute yase-Ethiopia yezempilo yomphakathi, uMnyango Wezempilo weFederal. Isu lesikhashana lesizwe kanye nesiqondisi sokuxilongwa kwelebhu yelebhu yeCovid-19. https://ephi.gov.et/mages/novel_coronavirus/ephi_phooc_covid-9_laboratoris_diagnosis_eng.pdf (Kufinyelelwe ku-20 Agasti 20) (EPHI, 2020).
UWolshin, S., Patel, N. & Keslelheim, njengoba ukuhlolwa okungalungile okungamanga kwe-SARS-Cov-2 Ukutheleleka Izingqinamba kanye nemiphumela. UWolshin, S., Patel, N. & Keslelheim, njengoba ukuhlolwa okungalungile okungamanga kwe-SARS-Cov-2 Ukutheleleka Izingqinamba kanye nemiphumela.UVoloshin S., Patel N. kanye neKesselheim njengezivivinyo ezingezinhle zamanga zeSARS-Cov-2 kanye nemiphumela yazo.UVolloshun S., Patel N. kanye neKesselheim njengezivivinyo ezingezinhle zokucasulwa kanye nomthelela wokutheleleka kwe-SARS-Cov-2. N. Eng. J. Medicine. 383 (6), e38 (2020).
I-MOULIUU, DS & Gourgoulianis, ama-KI amanga namanga namanga ama-covid-19 amacala we-19: amasu okuvimbela ukuphefumula kanye nokuphatha, ukugoma, nemibono eminye. I-MOULIUU, DS & Gourgoulianis, ama-KI amanga namanga namanga ama-covid-19 amacala we-19: amasu okuvimbela ukuphefumula kanye nokuphatha, ukugoma, nemibono eminye. I-MOULIUU, DS & Gourgoulianis, Ki ложжноте ложнноопожаительныеые Ibhola We-Okwamisa i-OPNELTY. I-MOULIUU, DS & Gourgoulianis, amacala amabi namanga amanga amabi we-Covid-19: amasu okuvimbela ukuphefumula kanye nokwelashwa, ukugoma kanye nendlela eya phambili.I-Muliu, DS kanye neGurgulianis, amacala amabi anamanga namanga angenamanga ama-Covid-19: amasu okuvimbela ukuphefumula nokwelashwa, ukugoma kanye nendlela eya phambili. Ukuphefumula uchwepheshe. umuthi. 15 (8), 993-1002 (2021).
I-MOULIUU, DS, iOannis, P. & Konstantinos, G. Ukuxilongwa kwe-Covil-19 emnyangweni wezimo eziphuthumayo: Ukubona lesihlahla kodwa kulahlekelwa ihlathi. I-MOULIUU, DS, iOannis, P. & Konstantinos, G. Ukuxilongwa kwe-Covil-19 emnyangweni wezimo eziphuthumayo: Ukubona lesihlahla kodwa kulahlekelwa ihlathi.I-Mouliou, DS, ioannis, P. noKonstantinos, G. Ukuxilongwa kwe-Covil-19 eMnyangweni Ophuthumayo: bheka isihlahla, ulahlekelwe yihlathi.I-Muliou DS, iOannis P., kanye ne-Konstantinos G. Covint-19 Ukuxilongwa kwamakamelo eziphuthumayo: Akekho amahlathi anele ezihlahla. Avele. umuthi. J. https:/doi.11366/12122219 (2022).
UDegli-Angeli, E. et al. Ukuqinisekiswa kanye nokuqinisekiswa kokusebenza kokuhlaziywa nokwelashwa kwe-abbott realtime realtime SARS-Cov-2 Assay. J. Clinical. Igciwane. 129, 104474. Https://doi.org/10.1016/J.JCV.2020.104474 (2020).
UMolliei, HR, Afshar, AA, Kalantar-Neyestanaki, D., FazlaliPour, M. UMolliei, HR, Afshar, AA, Kalantar-Neyestanaki, D., FazlaliPour, M. & Aflatoonia, B. Ukuqhathaniswa kwama-primer amahlanu okuqala kusuka ezifundeni ezihlukene zofuzo nge-RT-PCR evamile.UMolliei, HR, Afshar, AA, Kalantar-Neyestanaki, D., FazlaliPour, M. Ukuqhathaniswa kwama-aflatunyan, B. Ukuqhathaniswa kwama-genome amahlanu e-genome atholakala e-RT-PCR. UMolliei, HR, Afshar, AA, Kalantar-Neyestanaki, D., FazlaliPour, M. & Aflatoonia, B. 比较来自 CCR-19 不同基因组区域的五个引物组, 用于通过常规 RT-PRCR 检测病毒感染. UMolliei, HR, Afshar, AA, KALAntar-Neyestanaki, D., FazlaliPour, M. & Aflatoonia, B. Ukuqhathaniswa kwezindawo ezi-5 ezahlukahlukene zozakhi zofuzoI-Mollaei Hr, i-Afshar AA, Kalantar-Neyestanaki D, FazlaliPour M. Aflatunyan B. Ukuqhathaniswa kwamasethi amahlanu ama-primers avela ezifundeni ezahlukahlukene ze-genome evamile ye-RT-PCR.I-Iran. J. Microbiology. 12 (3), 185 (2020).
IGortrer, i. Et al. Imiphumela yokuqala yohlelo lukazwelonke lokuhlola ikhwalithi yangaphandle lokutholwa kwe-SARS-Cov-2 Genome Ukulandela ukulandelana. J. Clinical. Igciwane. 129, 104537. I-HTTPS: //doi.org/10.1016/J.JCV.2020.104537 (2020).
Wang, M. et al. Ukuhlolwa kokuhlaziywa kokusebenza kwempumelelo kwamakhithi amahlanu e-RT-PCR ye-Hlulis Huspiratory Syndrome coronavirus 2. J. Clinical. elebhu. anus. 35 (1), E23643 (2021).
Wang b. et al. Ukuhlaziywa kwezinhlelo eziyisikhombisa ezitholakala ngezentengiso ze-SARS-Cov-2 RNA Detection Kits eChina ngokusekelwe ekuphenduleni kwe-Poly-Time Chain Reaction real-time (PCR). Clinical. Amakhemikhali. elebhu. umuthi. 58 (9), E149-E153 (2020).
UVan Castersen, PB et al. Ukuqhathaniswa kwamakhithi ayisikhombisa we-RT-PCR-19 Diagnostic Kits. J. Clinical. Igciwane. 128, 104412 (2020).
Lu, yu, et al. Ukuqhathanisa ukusebenza kokuxilonga kwamakhithi amabili e-PCCR ngokutholwa kwe-SARS-Cov-2 Nucleic Acids. J. Clinical. elebhu. anus. 34 (10), E23554 (2020).
LeLLART, PR, njll. Ucwaningo lokuqhathanisa lwezinkundla zokuhlola ze-SARS ezine-cov-2 ze-nucleic-2 Ukuxilongwa. I-Microbiology. Ahluze. Diss. 99 (1), 115200 (2021).
I-abbott molecule. Abbott real-time sars-cov-2 ukuhlaziya iphakethe iphakheji Faka. I-https: //www.molecular.abbott/us/products/infectize-diise-disease/RealThe-sars-Cov-2-2-2-2-2-2-2--- 1-12. (Kusukela ngo-Agasti 10, 2020) (2020).
UKlein, S. et al. I-SARS-COV-2 RNA yokwahlukaniswa kwe-RNA isebenzisa ubuhlalu bamazibuthe bokutholwa okusheshayo okuphezulu nge-RT-QPCR ne-RT-Lamp. Igciwane 12 (8), 863 (2020).