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Mai ka laha ʻana o ka maʻi coronavirus (COVID-19) 2019, ua hoʻomohala ʻia ka nui o nā hoʻokolohua nucleic acid amplification tests (NAATs) a puni ka honua a ua lilo i mau hōʻike maʻamau.ʻOiai ua hoʻomohala koke ʻia kekahi mau hoʻokolohua a hoʻohana ʻia i nā hoʻokolohua diagnostic laboratory, ʻaʻole i loiloi ʻia ka hana o kēia mau hoʻokolohua ma nā ʻano hoʻonohonoho like ʻole.No laila, ua manaʻo kēia noiʻi e loiloi i ka hana o ka Abbott SARS-CoV-2, Daan Gene, BGI, a me Sansure Biotech assays me ka hoʻohana ʻana i ka Composite Reference Standard (CRS).Ua mālama ʻia ke aʻo ʻana ma ka Ethiopian Public Health Institute (EPHI) mai 1 a 30 Dekemaba 2020. Ua lawe ʻia nā 164 nasopharyngeal samples me ka hoʻohana ʻana i ka QIAamp RNA mini kit a me ka ʻōnaehana hoʻomākaukau hoʻohālike DNA Abbott.No 164 mau hōʻailona, 59.1% maikaʻi a 40.9% maikaʻi ʻole no CRS. Ua haʻahaʻa loa ka maikaʻi o Sansure Biotech i hoʻohālikelike ʻia me CRS (p <0.05). Ua haʻahaʻa loa ka maikaʻi o Sansure Biotech i hoʻohālikelike ʻia me CRS (p <0.05). Положительные результаты Sansure Biotech были значительно ниже по сравнению с CRS (p < 0,05). Ua emi loa nā hopena maikaʻi o Sansure Biotech i ka 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). He liʻiliʻi loa ka hopena maikaʻi o Sansure Biotech i hoʻohālikelike ʻia me CRS (p <0.05).ʻO ka ʻaelike holoʻokoʻa o nā loiloi ʻehā he 96.3-100% i hoʻohālikelike ʻia me CRS.Ma waho aʻe o ka haʻahaʻa haʻahaʻa haʻahaʻa o ka Sansure Biotech assay, ua aneane like ka hana o nā hoʻokolohua ʻehā.No laila, ʻo ka Sansure Biotech [Research Only (RUO)] assay e koi i ka hōʻoia hou no kona hoʻohana ʻana ma ʻAitiopa.ʻO ka hope, pono e noʻonoʻo ʻia nā noiʻi hou e loiloi i nā hoʻokolohua me nā ʻōlelo a ka mea hana kūpono.
ʻO ka hoʻokolohua Laboratory kahi ʻāpana o ka World Health Organization (WHO) Strategic Plan for Coronavirus Disease 2019 (COVID-19) Preparedness and Response (SPRP).Manaʻo ʻo WHO e pono nā ʻāina e kūkulu i ka hiki ke hoʻoikaika i ka mākaukau, ka hoʻokele hihia kūpono, ka makaʻala a me ka pane wikiwiki i nā pilikia olakino.Hōʻike kēia i ke kuleana o ka hale hana ke kī i ka ʻike ʻana i ka maʻi a me ka epidemiology o nā mea maʻi infectious e kū mai ana a me ka kāohi ʻana i kā lākou laha.
Pono ka hōʻailona o COVID-19 i ka ʻike maʻi maʻi a me ka ʻike olakino, nā hōʻailona pilikino/hōʻailona, a me ka ʻikepili radiographic a me ka ʻikepili2.Mai ka wā i hōʻike ʻia ai ka maʻi maʻi COVID-19 ma Wuhan, Kina, ua hoʻomohala ʻia ka nui o nā hoʻokolohua nucleic acid amplification tests (NAATs) a puni ka honua.Ua hoʻohana ʻia ka manawa maoli reverse transcription polymerase chain reaction (rRT-PCR) ma ke ʻano he hana maʻamau a maʻamau no ka hōʻoia ʻana o ka maʻi maʻi o ka maʻi hanu acute ʻeha 2 (SARS-CoV-2)3.Hoʻokumu ʻia ka ʻike molekole o SARS-CoV-2 i ka N (nucleocapsid protein gene), E (envelope protein gene), a me RdRp (RNA-dependent RNA polymerase gene) genes ma ORF1a/b (open reading frame 1a/b) .gene) wahi i ʻike ʻia mai ka genome viral.Ua manaʻo ʻia ʻo ia nā ʻāina mālama nui i loaʻa i nā genomes viral no ka ʻike ʻana i ka maʻi virus4.Ma waena o kēia mau genes, loaʻa i nā genes RdRp a me E ke kiʻekiʻe o ka ʻike analytical sensitivity, aʻo ka N genes he haʻahaʻa analytical sensitivity5.
Hiki ke ʻokoʻa ka hana ʻana o ka PCR assays ma muli o nā kumu like ʻole e like me: extraction reagents, amplification/detection reagents, extraction method, quality of the PCR machine and other instruments.Ma ʻApelila 2020, ʻoi aku ma mua o 48 mau mea diagnostic like ʻole mai ʻeiwa mau ʻāina i loaʻa i ka Emergency Use Authorization (EUA) no nā diagnostics COVID-196.Ma ʻAitiopia, ʻoi aku ma mua o 14 mau kahua PCR manawa maoli i hoʻohana ʻia no ka ʻike PCR o SARS-CoV-2 ma 26 mau keʻena olakino olakino, me ABI 7500, Abbott m2000, Roche 48000 a me Quant-studio7.Eia kekahi, loaʻa nā pahu hoʻāʻo PCR like ʻole, e like me ka ho'āʻo ʻo Daan Gene, ka hōʻike Abbott SARS-CoV-2, ka hoʻāʻo ʻo Sansure Biotech, a me ka hoʻāʻo ʻo SARS-CoV-2 BGI.ʻOiai paʻakikī loa ka rRT-PCR, hōʻike kekahi mau mea maʻi me COVID-19 i nā hopena maikaʻi ʻole ma muli o ka lawa ʻole o nā kope o ka viral ribonucleic acid (RNA) i nā laʻana ma muli o ka hōʻiliʻili pono ʻole, ka lawe ʻana, ka mālama ʻana a me ka mālama ʻana, a me ka hoʻāʻo ʻana i ka lab.nā kūlana a me nā hana a nā limahana8.Eia kekahi, ʻo ka laʻana a i ʻole ka hoʻokele mishandling, ka hoʻonohonoho ʻana i ka paepae pōʻai (Ct), a me ka cross-reactivity me nā ʻakika nucleic pathogenic a i ʻole ka SARS-CoV-2 RNA hiki ʻole ke alakaʻi i nā hopena maikaʻi ʻole i ka rRT-PCR9 assays.No laila, maopopo ka hiki i nā ho'āʻo PCR ke ʻike i nā mea lawe i nā ʻāpana gene, no ka mea ʻaʻole hiki iā lākou ke hoʻokaʻawale i waena o nā genes viral ikaika maoli, no laila hiki i nā hoʻokolohua ke ʻike wale i nā mea lawe a ʻaʻole nā mea maʻi10.No laila, he mea nui e loiloi i ka hana diagnostic me ka hoʻohana ʻana i nā ala maʻamau i kā mākou hoʻonohonoho.ʻOiai he nui nā NAAT reagents i loaʻa ma ka Ethiopian Public Health Institute (EPHI) a ma ka ʻāina holoʻokoʻa, ʻaʻole i hōʻike ʻia kahi loiloi hoʻohālikelike o ko lākou pono.No laila, ua manaʻo kēia noiʻi e loiloi i ka hana hoʻohālikelike o nā kits i loaʻa i ke kālepa no ka ʻike ʻana o SARS-CoV-2 e rRT-PCR me ka hoʻohana ʻana i nā specimens clinical.
ʻO ka huina o 164 mau mea komo me ka COVID-19 i manaʻo ʻia i komo i loko o kēia noiʻi.ʻO ka hapa nui o nā hōʻailona mai nā keʻena lapaʻau (118/164 = 72%), aʻo ke koena o 46 (28%) i komo mai nā kikowaena mālama ʻole.Ma waena o nā poʻe komo ʻaʻole i mālama ʻia ma ke kikowaena, 15 (9.1%) i hoʻopaʻa ʻia i nā hihia a me 31 (18.9%) i loaʻa i nā pilina o nā hihia i hoʻopaʻa ʻia.He kanaiwakūmākolu (56.7%) i komo i nā kāne, a ʻo ka mean (± SD) o ka poʻe i komo he 31.10 (± 11.82) mau makahiki.
Ma kēia haʻawina, ua hoʻoholo ʻia nā helu maikaʻi a maikaʻi ʻole o nā hoʻokolohua ʻehā no COVID-19.No laila, ʻo nā helu kūpono o ka assay Abbott SARS-CoV-2, Daan Gene 2019-nCoV assay, SARS-CoV-2 BGI assay, a me Sansure Biotech 2019-nCoV assay he 59.1%, 58.5%, 57.9% a me 55.5% kēlā me kēia. .He 97 (59.1%) a me 67 (40.9%) ka helu o ka ma'amau kuhikuhi (CRS).Ma kēia noiʻi ʻana, ua hoʻokumu ʻia ka wehewehe ʻana o CRS ma luna o ke kānāwai "kekahi maikaʻi", kahi o nā hopena hōʻike ʻehā, ʻelua a ʻoi aku paha nā hopena hōʻike i hāʻawi i ka hopena like i manaʻo ʻia he maikaʻi a maikaʻi ʻole paha.
Ma kēia haʻawina, ua loaʻa iā mākou kahi ʻaelike pākēneka maikaʻi ʻole (NPA) o 100% (95% CI 94.6–100) no nā loiloi āpau i hoʻohālikelike ʻia me CRS.Ua hōʻike ʻia ka loiloi Sansure Biotechnology i kahi PPA liʻiliʻi o 93.8% (95% CI 87.2-97.1) a me ka loiloi ʻo Daan Gene 2019-nCoV he ʻaelike holoʻokoʻa o 99.4% (95% CI 96.6-99.9).Ma ka hoʻohālikelike, ʻo ka ʻaelike holoʻokoʻa ma waena o ka SARS-CoV-2 BGI assay a me ka Sansure Biotech 2019-nCoV assay he 98.8% a me 96.3%, kēlā me kēia (Papa 2).
ʻO ka coefficient kappa o Cohen o ka ʻaelike ma waena o CRS a me Abbott SARS-CoV-2 i nā hopena assay i kūlike loa (K = 1.00).Pēlā nō, ʻike ʻia nā waiwai kappa a Cohen e Daan Gene 2019-nCoV, SARS-CoV-2 BGI, a me Sansure Biotech 2019-nCoV i kūlike pū me CRS (K ≥ 0.925).Ma kēia hoʻohālikelike hoʻohālikelike, ua hōʻike ka hōʻike chi-square (McNemar test) he ʻokoʻa loa nā hopena assay Sansure Biotech 2019-nCoV mai nā hopena CRS (p = 0.031) (Paukū 2).
E like me ka hōʻike ʻana ma ka Fig.1 ka pākēneka o ka waiwai Ct haʻahaʻa loa (<20 Ct) o Abbott SARS-CoV-2 assay (hui ʻia ʻo RdRp a me N gene) he 87.6% a me ORF1a/b gene Ct waiwai o Sansure Biotech 2019-nCoV assay i hōʻike i ka pākēneka o ka haʻahaʻa. ʻO ka waiwai Ct (<20 Ct) he 50.3% a ʻo ka waiwai Ct kiʻekiʻe (36-40 Ct) he 3.2%. 1 ka pākēneka o ka waiwai Ct haʻahaʻa loa (<20 Ct) o Abbott SARS-CoV-2 assay (hui ʻia ʻo RdRp a me N gene) he 87.6% a me ORF1a/b gene Ct waiwai o Sansure Biotech 2019-nCoV assay i hōʻike i ka pākēneka o ka haʻahaʻa. ʻO ka waiwai Ct (<20 Ct) he 50.3% a ʻo ka waiwai Ct kiʻekiʻe (36-40 Ct) he 3.2%.E like me ka hōʻike ʻana ma ka Fig.1, процент наименьшего значения Ct (< 20 Ct) анализа Abbott SARS-CoV-2 (комбинированный ген RdRp и N) составил 87,6%, а значение Ct гена ORF1a/b анализа Sansure Biotech 2019-nCoV показало что процент низкого значения Ct (< 20 Ct) составлял 50,3%, a высокое значение Ct (36–40 Ct) составляло 3,2%. 1, ʻo ka pākēneka o ka helu haʻahaʻa Ct haʻahaʻa (<20 Ct) ka nānā ʻana o Abbott SARS-CoV-2 (hui pū ʻia RdRp a me N) ʻo 87.6%, a ʻo ka Ct waiwai o ORF1a/b ʻano loiloi o Sansure Biotech 2019-nCoV i hōʻike ʻia. ʻo ka pākēneka o ka waiwai haʻahaʻa Ct (<20 Ct) he 50.3%, a he 3.2% ka waiwai kiʻekiʻe Ct (36-40 Ct).如 图рю值(< 20 Ct) 的百分比为50.3%，高Ct 值(36–40 Ct) 的百分比为3.2%. E like me ka mea i hōʻike ʻia ma ke Kiʻi 1, ʻo ka haʻahaʻa haʻahaʻa Ct waiwai pākēneka (<20 Ct) o Abbott SARS-CoV-2 hōʻike (hui pū ʻana o RdRp a me N gene) he 87.6%, ʻo ka ORF1a/b gene Ct waiwai o Sansure Biotech 2019-nCoV hōʻike. hōʻike haʻahaʻa Ct值(< 20 Ct) 的 pākēneka he 50.3%, 高Ct 值(36–40 Ct) 的 pākēneka he 3.2%. Как показаа на Та Та Та на konallah - Анализ nCoV показал низкий Ct. E like me ka mea i hōʻike ʻia ma ke Kiʻi 1, ʻo ka Abbott SARS-CoV-2 assay (ka hui pū ʻana i nā genes RdRp a me N) loaʻa ka haʻahaʻa haʻahaʻa haʻahaʻa Ct waiwai (<20 Ct) ma 87.6%, aʻo ka Ct waiwai o ka ORF1a/b gene i ka Sansure. ʻO ke aʻo ʻana o Biotech 2019 - Ua hōʻike ka loiloi o nCoV i kahi haʻahaʻa Ct. Процент значений (< 20 Ct) составил 50,3%, а процент высоких значений Ct (36–40 Ct) составил 3,2%. ʻO ka pākēneka o nā waiwai (<20 Ct) he 50.3%, a ʻo ka pākēneka o nā koina Ct kiʻekiʻe (36-40 Ct) he 3.2%.Ua hoʻopaʻa ʻia ka hōʻike Abbott SARS-CoV-2 B i nā waiwai Ct ma luna o 30. Ma ka ʻaoʻao ʻē aʻe, ma ka BGI SARS-CoV-2 assay ORF1a/b gene he kiʻekiʻe Ct waiwai (> 36 Ct) pākēneka he 4% (Fig. 1). Ma ka ʻaoʻao ʻē aʻe, ma ka BGI SARS-CoV-2 assay ORF1a/b gene he kiʻekiʻe Ct waiwai (> 36 Ct) pākēneka he 4% (Fig. 1). С другой стороны, в анализе BGI SARS-CoV-2 ген ORF1a/b имел высокое значение Ct (> 36 Ct), процент которого составл. 1%). Ma ka ʻaoʻao ʻē aʻe, i ka nānā ʻana o BGI SARS-CoV-2 gene ORF1a/b i loaʻa kahi waiwai Ct kiʻekiʻe (> 36 Ct), ʻo ka pākēneka he 4% (Fig. 1).另一方面，在BGI SARS-CoV-2 检测中，ORF1a/b 基因具有高Ct 伈> 36 Ct）的百分比为4%（图 Ma ka ʻaoʻao ʻē aʻe, ma ka BGI SARS-CoV-2 ʻike, ʻo ka pākēneka o ORF1a/b gene me ka waiwai Ct kiʻekiʻe (>36 Ct) ʻo 4% (Figure 1). С другой стороны, в анализе BGI SARS-CoV-2 процент генов ORF1a/b с высокими значениями Ct (>36 Ct) составил 4% (e like me 1). Ma ka ʻaoʻao ʻē aʻe, ma ka loiloi BGI SARS-CoV-2, ʻo ka pākēneka o nā genes ORF1a/b me nā koina Ct kiʻekiʻe (>36 Ct) he 4% (Fig. 1).
I loko o kēia haʻawina, ua lawe mākou i 164 nasopharyngeal samples.No nā ʻano hoʻokolohua āpau, ua hoʻokaʻawale ʻia ka RNA a me ka hoʻonui ʻana me ka hoʻohana ʻana i nā ʻano a me nā kits i ʻōlelo ʻia e nā mea hana.
Ua hōʻike ʻia kēia haʻawina ʻo ka hoʻāʻo ʻana o Abbott no SARS-CoV-2 i ka hana ʻike like me CRS, me ka 100% maikaʻi, maikaʻi ʻole, a me ka concordance holoʻokoʻa.ʻO ka aelike kappa a Cohen he 1.00, e hōʻike ana i ka ʻaelike piha me CRS.Ua ʻike ʻia kahi noiʻi like ʻana e ke Kulanui o Wakinekona ma US ʻo ka ʻike holoʻokoʻa a me ka kikoʻī o ka hoʻāʻo Abbott no SARS-CoV-2 he 93% a me 100%, i hoʻohālikelike ʻia i ka hoʻoholo ʻana i ka lab (LDA) o ka CDC. .11. Hoʻokumu ʻia ka ʻōnaehana ʻike Abbott SARS-CoV-2 ma ka ʻike hui pū ʻana o nā genes N a me RdRp, no ka mea, ʻoi aku ka maʻalahi o nā genes ʻelua, e hōʻemi ana i nā mea ʻino ʻole12.Ua hōʻike pū kekahi haʻawina ma Vienna, Austria, ua hōʻemi ʻia nā hopena hoʻoheheʻe a me ka hoʻonui ʻana i ka pono o ka ʻike.No laila, hiki ke hoʻopili ʻia ka hoʻokūkū kūpono o Abbott no ka SARS-CoV-2 assay me kahi ʻōnaehana ʻike platform e ʻike pū ana i nā genes combinatorial, unuhi i ka nui o nā laʻana (0.5 ml), a hoʻohana i ka nui o ka eluent (40 µl).
Ua hōʻike pū ʻia kā mākou hopena i ka hana ʻike ʻana o ka hoʻāʻo genetic Daan ua aneane like me ka CRS.Ua kūlike kēia me kahi haʻawina14 i mālama ʻia ma ke Kulanui ʻo Anhui ma Huainan, Kina, a me ka ʻōlelo a ka mea hana no 100% ʻaelike maikaʻi.ʻOiai ka hōʻike ʻana o nā hopena like ʻole, ua hewa kekahi laʻana ma hope o ka hoʻāʻo hou ʻana i ka eluate like, akā maikaʻi ma ka Abbott SARS-CoV-2 a me Sansure Biotech nCoV-2019 assays.Hōʻike kēia i nā ʻano like ʻole o nā hopena ma nā ʻano hoʻokolohua like ʻole. Eia nō naʻe, i loko o ke aʻo ʻana i hana ʻia ma Kina15, ʻokoʻa loa ka hopena o ka ho'āʻo ʻo Daan Gene (p <0.05) i hoʻohālikelike ʻia i kā lākou hōʻike kuhikuhi i wehewehe ʻia e ka lab. Eia nō naʻe, i loko o ke aʻo ʻana i hana ʻia ma Kina15, ʻokoʻa loa ka hopena o ka ho'āʻo ʻo Daan Gene (p <0.05) i hoʻohālikelike ʻia i kā lākou hōʻike kuhikuhi i wehewehe ʻia e ka lab. Тем не менее, в исследовании, проведенном в Китае15, результат анализа Daan Gene значительно отличалсято (p < 0,05) лато (p < 0,05) лит Eia nō naʻe, i loko o kahi noiʻi ma Kina15, ʻokoʻa loa ka hopena o ka hōʻike ʻana a Daan Gene (p <0.05) mai kā lākou hōʻike hōʻike hōʻike.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义的参考检测看0.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定义的参考检浔盠检测看. Однако в исследовании, проведенном в Китае15, результаты генетического теста Daan значительно отличалисьно отличались (p молюнось) < 0,000 Eia naʻe, ma kahi noiʻi ma Kina15, ʻokoʻa loa nā hopena o ka hoʻāʻo genetic a Daan (p <0.05) i hoʻohālikelike ʻia me kāna hōʻike hoʻokolohua hōʻike.ʻO kēia ʻokoʻa paha ma muli o ka naʻau o ka hōʻike kuhikuhi e ʻike ai iā SARS-CoV-2, a he mea nui paha nā haʻawina hou e hoʻoholo ai i ke kumu.
Eia kekahi, ua loiloi kā mākou noiʻi ʻana i ka hana hoʻohālikelike o ka SARS-CoV-2 BGI assay me CRS, e hōʻike ana i ka ʻaelike pākēneka maikaʻi maikaʻi loa (PPA = 97.9%), ka ʻaelike pākēneka maikaʻi ʻole (NPA = 100%), a me ka ʻaelike pākēneka holoʻokoʻa e ke kāne ( OPA).).= 98.8%).Ua hōʻike maikaʻi nā waiwai Kappa a Cohen (K = 0.975).Ua hōʻike nā haʻawina ma Netherlands16 a me Kina15 i nā hopena kūlike.ʻO ka hōʻike ʻo SARS-CoV-2 BGI kahi hōʻike ʻike hoʻokahi gene (ORF1a/b) me ka hoʻohana ʻana i 10 µl amplification/detection eluate.ʻOiai ʻo ka ʻaelike helu helu maikaʻi me kā mākou mau hualoaʻa, ua hala ʻelua mau laʻana maikaʻi (1.22%) o ka hōʻailona holoʻokoʻa.Hiki i kēia ke loaʻa i nā hopena lapaʻau nui no ka hoʻoili ʻana i ka dinamika ma ka pae maʻi a me ke kaiāulu.
ʻO kahi loiloi hoʻohālikelike ʻē aʻe i komo i loko o kēia haʻawina ʻo ka Sansure Biotech nCoV-2019 rRT-PCR (RUO) assay;ʻo ka pākēneka pāʻani holoʻokoʻa he 96.3%.Ua hoʻoholo pū ʻia ka ikaika o ka ʻaelike e ka Cohen's Kappa waiwai, ʻo ia ka 0.925, e hōʻike ana i ka ʻaelike piha me ka CRS.Eia hou, ua like kā mākou hopena me nā haʻawina i mālama ʻia ma Central South University ma Changsha, Kina, a ma ke Keʻena Laboratory Clinical o Liuzhou People's Hospital, Liuzhou City, Kina17. ʻOiai ua hoʻopaʻa ʻia ka concordance helu maikaʻi ma luna, ua hōʻike ka hōʻike chi-square (MacNemar test) i ka hopena o ka Sansure Biotech assay he ʻokoʻa koʻikoʻi koʻikoʻi ke hoʻohālikelike ʻia me CRS (p <0.005). ʻOiai ua hoʻopaʻa ʻia ka concordance helu maikaʻi ma luna, ua hōʻike ka hōʻike chi-square (MacNemar test) i ka hopena o ka Sansure Biotech assay he ʻokoʻa koʻikoʻi koʻikoʻi ke hoʻohālikelike ʻia me CRS (p <0.005). Несмотря на то, что было зафиксировано указанное выше хорошее статистическое соответствие, критерий хи-квадрат (критерий Макнемара) показал, что результат анализа Sansure Biotech имеет статистически значимое различие по сравнению с CRS (p < 0,005). ʻOiai ua hoʻopaʻa ʻia ka ʻaelike helu helu maikaʻi ma luna, ua hōʻike ka hōʻike chi-square (McNemar test) i ka hopena o ka Sansure Biotech assay he ʻokoʻa koʻikoʻi koʻikoʻi ke hoʻohālikelike ʻia me ka CRS (p <0.005).尽管 记录 述 述述 a 良好 的 致性 viema ;: Ala-尽管尽管 致了 致良好 性性 性性 性性 致性 致性 (Malinuma 检验检验, Hono-(P <0.005. …….）））） На на на Иот ххротиларатиий крнтачим павтаким пк <0 0 0, 0,25) Кр соказачим паои ʻOiai ka ʻaelike helu helu maikaʻi i hōʻike ʻia ma luna, ua hōʻike ka hōʻike chi-square (McNemar test) i kahi ʻokoʻa koʻikoʻi (p <0.005) ma waena o ka Sansure Biotech assay a me ka CRS.ʻEono mau laʻana (3.66%) i ʻike ʻia he mau hewa wahaheʻe i hoʻohālikelike ʻia me CRS (Supplementary Table 1);He mea koʻikoʻi kēia, ʻoi aku ka nui o ka hoʻoili ʻana o ka maʻi maʻi.Kākoʻo ka ʻikepili i luna i kēia helu haʻahaʻa haʻahaʻa15.
Ma kēia haʻawina, ua hoʻoholo ʻia nā waiwai Ct no kēlā me kēia assay a me kēlā me kēia kahua, me ka haʻahaʻa haʻahaʻa haʻahaʻa Ct waiwai i hōʻike ʻia ma ka Abbott SARS-CoV-2 assay.Pili paha kēia hopena i ka ʻōnaehana hoʻāʻo genetic hui pū ʻana o Abbott no ka ʻike ʻana o SARS-CoV-2.No laila, e like me ka Figure 1, 87.6% o nā hopena Abbott SARS-CoV-2 i loaʻa nā waiwai Ct ma lalo o 20. He helu liʻiliʻi wale nō o nā hopena hoʻohālike (12.4%) i ka pae 20-30.ʻAʻole i hoʻopaʻa ʻia nā waiwai Ct ma luna o 30.Ma kahi o ka hoʻohana ʻana o Abbott i ka ʻano hoʻāʻo genetic panel SARS-CoV-2, pili paha kēia hopena i ka palena haʻahaʻa haʻahaʻa (32.5 RNA kope/mL)18, ʻekolu mau manawa haʻahaʻa ma mua o ka palena haʻahaʻa o ka hui o 100 RNA kope. /mL.ml)19.
He mau palena ko kēia haʻawina: ʻo ka mea mua, ʻaʻohe o mākou kaʻina hana maʻamau/pili [e like me ka viral load a i ʻole nā hoʻokolohua hoʻokolohua ʻē aʻe (LDA)] ma muli o ka nele o nā kumuwaiwai.ʻO ka lua, ʻo nā hiʻohiʻona a pau i hoʻohana ʻia i kēia noiʻi ʻana he mau nasopharyngeal swabs, ʻoiai ʻaʻole pili nā hopena i nā ʻano specimen ʻē aʻe, a ʻo ke kolu, he liʻiliʻi kā mākou hāpana.
Ua hoʻohālikelike kēia haʻawina i ka hana o ʻehā rRT-PCR assays no SARS-CoV-2 me ka hoʻohana ʻana i nā laʻana nasopharyngeal.ʻO nā hoʻokolohua ʻike a pau ua aneane like ka hana, koe wale ka Sansure Biotech assay. Ma waho aʻe, ua ʻike ʻia ka helu haʻahaʻa haʻahaʻa i ka Sansure Biotech assay i hoʻohālikelike ʻia me ka CRS (p <0.05). Ma waho aʻe, ua ʻike ʻia ka helu haʻahaʻa haʻahaʻa i ka Sansure Biotech assay i hoʻohālikelike ʻia me ka CRS (p <0.05). Кроме того, в тесте Sansure Biotech был выявлен низкий процент положительных результатов по сравнению с CRS (p < 0,05). Eia kekahi, ua hōʻike ka hōʻike ʻo Sansure Biotech i kahi haʻahaʻa haʻahaʻa o nā hopena maikaʻi i hoʻohālikelike ʻia me 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). Eia kekahi, ʻo ka Sansure Biotech assay he haʻahaʻa haʻahaʻa haʻahaʻa i hoʻohālikelike ʻia me CRS (p <0.05).ʻO ka loiloi Sansure Biotech nCoV-2019 (RUO) o PPA, NPA a me ka ʻaelike holoʻokoʻa i ʻoi aku ma mua o 93.5% me kahi Cohen Kappa ikaika o ka ʻaelike waiwai o 0.925.ʻO ka hope, pono ka Sansure Biotech Assay (RUO) i ka hōʻoia hou ʻana no ka hoʻohana ʻana ma ʻAitiopa, a pono e noʻonoʻo ʻia nā noiʻi hou e loiloi i nā koi mai nā mea hana pilikino.
Ua mālama ʻia ka hoʻolālā haʻawina hoʻohālikelike ma nā keʻena olakino ʻehā ma Addis Ababa, ka Halemaʻi ʻo Eka Kotebe, ka Halemai Millennium Church Treatment Center, Zewooditu Memorial Hospital, a me St. Peter's Tuberculosis Specialist Hospital.Ua hōʻiliʻili ʻia ka ʻikepili ma waena o Dekemaba 1 a me 31, 2020. Ua koho pono ʻia nā keʻena olakino no kēia noiʻi ma muli o kā lākou helu kiʻekiʻe o nā hihia a me ka loaʻa ʻana o nā kikowaena lapaʻau nui ma ke kūlanakauhale.Pēlā nō, ua koho ʻia nā mea hana, me ka ABI 7500 a me Abbott m2000 PCR pono manawa, e like me nā ʻōlelo aʻoaʻo a nā mea hana reagent NAAT, a ua koho ʻia nā pahu ʻike PCR ʻehā no kēia haʻawina, ʻoiai ua hoʻohana ka hapa nui o nā laboratories ma ʻAitiopa ma ka liʻiliʻi loa. ʻehā o lākou.Hōʻike Gene, hōʻike Abbott SARS-CoV-2, hōʻike ʻo Sansure Biotech, a me SARS-CoV-2 BGI hōʻike i hana ʻia i ka wā o ke aʻo ʻana).
Ua hana ʻia ka hoʻāʻo ʻana no SARS-CoV-2 mai 1 a 30 Dekemaba 2020 me ka hoʻohana ʻana i 3 ml o Viral Transport Medium (VTM) (Miraclean Technology, Shenzhen, Kina) mai nā poʻe i hoʻokolokolo ʻia no COVID-19 i kuhikuhi ʻia iā EPHI.Ua hōʻiliʻili ʻia nā laʻana nasopharyngeal e nā mea hōʻiliʻili laʻana i aʻo ʻia a hoʻouna ʻia iā EPHI i loko o nā ʻeke ʻekolu.Ma mua o ka hoʻokaʻawale ʻana i ka waikawa nucleic, hāʻawi ʻia kēlā me kēia laʻana i kahi helu ʻike kūʻokoʻa.Hana ʻia ka unuhi ʻana mai kēlā me kēia laʻana i ka wā e hōʻea ai me ka hoʻohana ʻana i nā ʻano hoʻoheheʻe manual a me ka ʻakomi.No laila, no ka 'akomi extraction o Abbott m2000, 1.3 ml (me 0.8 ml make leo a me 0.5 ml extraction inlet leo) o ka hāpana i extracted mai kēlā me kēia hāpana a hele aku i loko o ka Abbott DNA Sample Preparation System (Abbott Molecular Inc. des Plaines. IL, USA).) Ua hoʻokomo ʻia kahi pūʻulu o 96 [92 laʻana, ʻelua mau mana ʻike a me ʻelua mau mana non-template (NTC)] i loko o ke kaʻina holoʻokoʻa (hoʻihoʻi a ʻike) ʻelua pōʻai o SARS-CoV-2 (EUA) i ka manawa maoli.eli ana.Pēlā nō, no ka unuhi lima, e hoʻohana i nā laʻana like (no ka unuhi ʻakomi a me ka ʻike).No laila, i loko o ke kaʻina hana, 140 µl laʻana i aliquoted a unuhi ʻia me ka QIAamp Viral RNA Mini Kit (QIAGEN GmbH, Hilden, Kelemānia) i nā pūʻulu o 24 (me 20 mau laʻana, ʻelua mana assay a me ʻelua NTC) ma ʻeiwa mau pōʻai.Ua hoʻonui ʻia a ʻike ʻia nā ʻeluates i unuhi ʻia me ka hoʻohana ʻana i kahi mea hoʻokele wela ABI 7500 e hoʻohana ana i ka SARS-CoV-2 BGI assay, Daan Gene assay, a me Sansure Biotech assay.
ʻO ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana o SARS-CoV-2 viral RNA e hahai ana i ka loina bead magnetic me ka hoʻohana ʻana i nā reagents hoʻomākaukau hoʻomākaukau ʻana o Abbott DNA.Hoʻohana ʻia ka inactivation o nā laʻana a me ka solubilization o nā ʻāpana viral me ka hoʻohana ʻana i kahi mea holoi i loaʻa ka guanidine isothiocyanate e hoʻopau i ka protein a hoʻopau i ka RNase.A laila hoʻokaʻawale ʻia ka RNA mai ka protein ma o ka hoʻokaʻawale ʻana i ka māhele paʻa me ka silica, ʻo ia hoʻi ka paʻakai guanidinium a me ka pH alkaline o ka lysis buffer e hoʻoikaika i ka hoʻopaʻa ʻana o nā waikawa nucleic i ke silica (SiO2).Hoʻopau ke kaʻina hoʻomaʻemaʻe i ke koena o nā protein a me nā ʻōpala e hana i kahi hopena maʻemaʻe.Hoʻokaʻawale ʻia ʻo RNA maʻemaʻe mai nā microparticles e pili ana i ka silica me ka hoʻohana ʻana i ke kahua magnetic o ka mea kani20,21.Ma ka ʻaoʻao ʻē aʻe, lawe ʻia ka hoʻokaʻawale lima a me ka hoʻomaʻemaʻe ʻana i ka RNA e ke ʻano kolamu spin me ka hoʻohana ʻana i ka centrifugation ma kahi o kahi kū magnetic a me ka hoʻokaʻawale ʻana o nā microparticles mai ka eluent.
Ua hana ʻia ka Abbott Real-Time SARS-CoV-2 Detection Test (Abbott Molecular, Inc.) e like me nā ʻōlelo a ka mea hana, i loaʻa iā EUA19,22 mai ka WHO a me FDA.Ma kēia kaʻina hana, ua hana ʻia ka inactivation laʻana ma mua o ka unuhi ʻana i ka ʻauʻau wai ma 56 ° C no 30 min.Ma hope o ka inactivation virus, ua hana 'ia ka nucleic acid extraction ma ka Abbott m2000 SP mea kani mai 0.5 ml VTM me ka Abbott m2000 DNA sample preparation system.e like me ka mea hana.Hana ʻia ka hoʻonui a me ka ʻike ʻana me ka Abbott m2000 RT-PCR, a ua hana ʻia ka ʻike ʻelua no nā genes RdRp a me N.ROX) a me VIC P (proprietary dye) no ka huli ʻana a me ka ʻike ʻana i nā mana o loko, e ʻae ana i ka ʻike like ʻana o nā huahana amplification ʻelua 19 .
Hoʻokumu ʻia ke ʻano ʻike hoʻonui o kēia kit ma kahi ʻenehana RT-PCR hoʻokahi.Ua koho ʻia nā genes ORF1a/b a me N ma ke ʻano he ʻāina mālama ʻia e Daan Gene Technology e ʻike i ka hoʻonui ʻia ʻana o ka ʻāina.Ua hoʻolālā ʻia nā kumu mua a me nā ʻimi fluorescent (N gene probes i kapa ʻia me FAM, ORF1a/b probes i hōʻailona ʻia me VIC) e ʻike i ka SARS-CoV-2 RNA i nā laʻana.Ua hoʻomākaukau ʻia ka hui ʻana o ka eluent a me ka master ma ka hoʻohui ʻana i 5 µl o ka eluent i 20 µl o ka hui master i ka nui hope o 25 µl.Hana ʻia ka hoʻonui ʻana a me ka ʻike ʻana ma kahi mea hana PCR manawa maoli ABI 750024.
Ua ʻike ʻia nā genes ORF1a/b a me N me ka hoʻohana ʻana i ka Sansure Biotech nCoV-2019 Nucleic Acid Diagnostic Kit (fluorescent PCR detection).E hoʻomākaukau i nā ʻimi kikoʻī no kēlā me kēia moʻo i manaʻo ʻia ma ke koho ʻana i ke kahawai FAM no ka ʻāpana ORF1a/b a me ke kahawai ROX no ka gene N.I kēia pahu hoʻāʻo, ua hoʻohui ʻia nā mea hoʻoheheʻe eluent a me master mix reagent penei: hoʻomākaukau i 30 µl master mix reagent a me 20 µl eluted sample no ka ʻike/hoʻonui.Ua hoʻohana ʻia ʻo PCR ABI 750025 manawa maoli no ka hoʻonui ʻana/ʻike.
ʻO ka hōʻike ʻo SARS-CoV-2 BGI kahi pahu rRT-PCR manawa maoli no ka maʻi maʻi o COVID-19.Aia ka ʻāina i hoʻopaʻa ʻia ma ka ʻāpana ORF1a/b o ka genome SARS-CoV-2, ʻo ia ke ʻano ʻike ʻano hoʻokahi.Eia kekahi, ʻo ka gene β-actin hale mālama kanaka he gene i hoʻoponopono ʻia i loko.Hoʻomākaukau ʻia ka master mix ma ka hui ʻana i 20 µl o ka master mix reagent a me 10 µl o ka laʻana RNA i unuhi ʻia i loko o kahi pā lua26.Ua hoʻohana ʻia kahi mea hana PCR manawa maoli ABI 7500 fluorescent quantitative no ka hoʻonui a me ka ʻike.ʻO ka hoʻonui ʻana i ka waikawa nucleic a pau, nā kūlana holo PCR no kēlā me kēia assay, a me ka wehewehe ʻana i nā hopena i hana ʻia e like me nā ʻōlelo a ka mea hana (Papa 3).
Ma kēia hoʻohālikelike hoʻohālikelike, ʻaʻole mākou i hoʻohana i ke ʻano maʻamau kuhikuhi no ka hoʻoholo ʻana i ka pākēneka o ka ʻaelike (maikaʻi, maikaʻi ʻole, a holoʻokoʻa) a me nā ʻāpana hoʻohālikelike ʻē aʻe no nā loiloi ʻehā.Ua hana ʻia kēlā me kēia hoʻohālikelike hoʻāʻo me CRS, ma kēia aʻo ʻana ua hoʻonohonoho ʻia ka CRS e ke kānāwai "kekahi maikaʻi" a ua hoʻoholo ʻia ka hopena, ʻaʻole ma ka hoʻāʻo hoʻokahi, ua hoʻohana mākou i ʻelua mau hopena hoʻokolohua like ʻole.Eia kekahi, i ka hihia o ka hoʻouna ʻana o COVID-19, ʻoi aku ka pōʻino o nā hopena maikaʻi ʻole ma mua o nā hopena maikaʻi ʻole.No laila, no ka ʻōlelo ʻana i ka "maikaʻi" e like me ka pololei mai kahi hopena CRS, pono ʻelua mau ho'āʻo ho'āʻo ʻoi aku ka maikaʻi, ʻo ia hoʻi, ʻo ka liʻiliʻi loa e loaʻa mai ana hoʻokahi hopena maikaʻi mai kahi hōʻike EUA.No laila, mai loko mai o ʻehā mau hopena hoʻāʻo, ʻelua a ʻoi aku paha nā hopena hōʻike e hāʻawi i ka hopena like i manaʻo ʻia he maikaʻi a maikaʻi ʻole paha18,27.
Ua hōʻiliʻili ʻia ka ʻikepili me ka hoʻohana ʻana i nā palapala unuhi ʻikepili i kūkulu ʻia, hoʻokomo ʻia ka ʻikepili a me ka nānā ʻana me ka polokalamu helu helu Excel a me SPSS version 23.0 no nā ʻikepili wehewehe.Ua kālailai ʻia ka ʻaelike maikaʻi, ʻino, a me ka hapa nui, a ua hoʻohana ʻia kahi helu Kappa e hoʻoholo ai i ke degere o ka ʻaelike o kēlā me kēia ʻano me CRS.Ua unuhi ʻia nā waiwai Kappa penei: 0.01 a 0.20 no ka ʻaelike haʻahaʻa, 0.21 a 0.40 no ka ʻaelike maʻamau, 0.41-0.60 no ka ʻaelike kūpono, 0.61-0.80 no ka ʻaelike nui a me 0.81-0.99 no ka ʻaelike piha28.
Ua loaʻa mai ka ʻae kūpono mai ke Kulanui o Addis Ababa a ua ʻae ʻia nā protocol hoʻokolohua āpau no kēia noiʻi ʻana e ka Papa Hoʻonaʻauao ʻepekema ʻepekema ʻo Ethiopian Public Health Institute.ʻO ka helu kuhikuhi no ka EPHI Ethics License ʻo EPHI/IRB-279-2020.Ua hoʻohana ʻia nā ʻano hana āpau e like me nā ʻōlelo aʻoaʻo a me nā ʻōlelo a ka ʻAitiopia National Comprehensive Guidelines no ka mālama ʻana o COVID-19.Eia hou, ua loaʻa ka ʻae ʻike i kākau ʻia mai nā mea aʻo a pau ma mua o ke komo ʻana i ka haʻawina.
Aia nā ʻikepili a pau i loaʻa a i ʻike ʻia i loko o kēia haʻawina i loko o kēia ʻatikala i paʻi ʻia.Loaʻa nā ʻikepili e kākoʻo ana i nā hopena o kēia noiʻi mai ka mea kākau ma muli o ke noi kūpono.
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Ka manawa hoʻouna: Dec-08-2022