Perficientur quattuor nucleic acidum amplificationem assays ad identify Sars, Cov- II in Aethiopia

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Cum MMXIX Coronavirus morbus (CoviD-XIX), tumultuarios, multi commercial nucleic acidum amplificationem probat (Naats) sunt developed circum orbem terrarum et facti sunt vexillum assays. Licet pluribus probat cito developed et applicantur ad laboratorium Diagnostic probat, in his probat probat non aestimari in varietate occasus. Ideo hoc studium intendebant ad aestimandas perficientur abbott Sars-Cov-II, Daan Gene, BGI et Sanseure Biotech usura composito referat Latin (CS). In studiis erat conducted ad Aethiops Public Health Institutum (Ephi) ex I ad XXX Decembris MMXXX. CLXIV Nasopharyngeos exempla et Abbott Using in Qiaamp RNA Mini Ornamentum et Abbott DNA Sample Praeparatio. De CLXIV specimens, 59.1% erant positivum et 40,9% fuerunt negative pro CRS. Sansure Biotech positivity erat significantly humilis comparari CRS (P <0.05). Sansure Biotech positivity erat significantly humilis comparari CRS (P <0.05). Положительные результаты Sansure Biotech были значительно Ниже по Сравнению с Crs (P <0,05). Sansure Biotech scriptor positivum eventus erant significantly inferior comparari CRS (P <0.05).与 CS 相比, Sansure Biotech 的阳性率显着较低 (P <0.05).与 CS 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 cum significantly paucioribus positivum results comparari CRS (P <0.05).In altiore consensu quattuor analyses erat 96.3-100% comparari CRS. In addition ad humilis positivity rate de sansure biotech asay, in perficientur de quattuor assays erat prope comparabiles. Ut talis, in Sansure Biotech [Research Tantum (Ruo)] Assous exigit additional sanatio ad usum in Aethiopia. Denique additional investigationis debet considerari ad aestimandas assays cum oportet manufacturer scriptor petat.
Laboratory testing est pars mundi salutem organization (qui) opportuna Plan ad coronavirus morbus MMXIX (Covi, XIX) praeparatio et responsio (Sprop). Qui monet, quod regiones postulo aedificare elit facultatem ad amplio praeparatio, propriis casu procuratio, vigilantia et celeri responsio ad publicam salutem challenges. Hoc insinuat quod partes de officinarum est clavis ad characterizing morbus et epidemiology of emergentes infectiosis agentibus et moderante propagationem.
Et diagnosis de Covi, XIX requirit epidemologicas et medicinae notitia, personalis symptoms / signa et radiographic et laboratorium data2. Cum autem Covi-XIX tum nuntiavit in Wuhan, Sina, multi commercial nucleic acidum amplificationem probat (Naats) sunt developed circum orbem terrarum. Real-vicis vicissim transcriptionem Polymerase catena reactionem (RRT-PCR) est usus ut a exercitatione et vexillum modum ad laboratorium diagnosis gravi acuti respiratoriorum syndrome II (Sars-II-II) III infectio. Molecular Deprehensio Sars, Cov-II est typically secundum N (nucleocapsid protein gene), e (RNA dapibus gene), et rna / b (Open Lectio frame 1a / b). gene) regionem identified ex virales. Cogita esse principalis conserventur regiones in virales genomes virum recognition4. Inter haec genes, et rdrp et E genes habere altum analytica deprehendatur sensibilitatem, cum n gene est humilis analyticae sensitivity5.
In perficientur PCR assays potest variantur fretus variis factoribus ut: extraction Reagents, amplificationem / Deprehensio Reagents, extraction modum, qualis est PCR apparatus et aliarum instrumentis. Sicut Aprilis MMXX, magis quam XLVIII diversis diagnostic cogitationes de novem terris non receperunt emergency usu LICENTIA (EUA) ad CXCVI CXCVI Diagnostics. In Aethiopia, magis quam XIV Real-vicis PCR Platforms sunt pro PCR Deprehensio Sars-Cov-II ad XXVI Public Health Institutionum, inter DCCV (VII) D, Abbott M2000, Roche (XLV) et Quant-Studio7. In addition, variis PCR test kits sunt available, ut Daan Gene test, Abbott Sars, Cov-II test, sansure BIOTECH test, et Sars-Cov-II BGI test. Licet RRT-PCR est altus sensitivo, quidam aegris cum Covi-XIX Report falsum negative eventus debitum ad insufficiens exemplaria virales citrum acidum (RNA) in exemplaria ex improprium, onerariis. Condiciones et actionibus ex personnel8. In addition, sample aut potestate mishandling, exolvuntur limine (CT) occasum, et crucem-reactivity cum aliis morbificae nucleic acida aut inertes / RELICTUM Sars-COV-II RNS potest ad falsum positivum results in RRN, PCR9 ADSYS. Ut patet quod PCR probat potest quidem identify carriers de gene fragmenta, ut non distinguere inter vere activa virales genes, ita probat non solum identify carriers et non a patientiam. Ideo est momenti ad assess Diagnostic perficientur per vexillum modi in occasum. Licet plures naat reagentia sunt praesto ad Aethiopiam publicam salutem Institutum (Ephi) et in regione, nihil comparativum aestimatione eorum efficaciam habet tamen nuntiavit. Unde hoc studium intendebant ad aestimandas comparativum perficientur commercially available Kits pro deprehendatur Sars-Cov- II by RRT-PCR usura orci specimens.
A summa CLXIV participantium cum suspected Covi, XIX sunt includitur in hoc studium. Plures ex exemplaria ex curatio centers (118/164 = LXXII%), cum reliqua XLVI (XXVIII%) participantium fuerunt ex non-curatio centers. Inter participantium non agitur ad centrum, XV (9.1%) cum esset clinically suspected casibus et XXXI (18.9%) cum contactus confirmata casibus. Nonaginta tres (56.7%) participantium fuerunt masculum et medium (± s) aetatis participantium erat 31.10 (± 11.82) annis.
In hoc studiis, positivum et negans rates quattuor probat pro Covi XIX et determinari. Ut, positivum rates de abbott Sars, Cov-II asay, Daan Gene MMXXIX-NCOV Assay, Sars-Cov-II BGI Assay, et Sansure BIOTECH-NCOV erant 59.1%, 58.5%, 57.9 et 58.5% et 58.5. Et positivum et negative compositum referat vexillum (CRS) scores fuerunt XCVII (59.1%) et LXVII (40.9%), respectively (mensa I). In hoc studium, quod definitio CRS fundatur in "quis positivum" regula, quibus ex quatuor test results, duo vel plures test eventus, qui dedit idem effectus habentur vera positive vel negativa.
In hoc studio, invenimus negans recipis pactum (NPA) de C% (XCV% CI 94.6-100) omnes analys comparari CRS. In Sansure Analysis ostendit a minimam PPA de 93.8% (XCV% CI 87.2-97.1) et Daan Gene MMXXIX-NCOV Analysis habebat in altiore consensu 99,4% (XCV% CI 96.6-99.9 (XCV% CI 96.6-99.9 (XCV% CI 96.6-99.9). In Contra, altiore consensu inter Sars-Cov-II BGI Assay et Sansure Biotech MMXIX-NCOV erat 98.8% et 96.3%, respective (Table II).
Cohen scriptor Kappa coefficiente pactum inter CRS et abbott Sars, Cov- II Assay results erat plene consistent (k = 1.00). Similiter, Cohen scriptor Kappa values ​​deprehendi a Daan Gene MMXIX, NCOV, Sars, Cov-II BGI, et Sansure Biotech MMXIX, NCOV sunt etiam plene consistent cum CRS (K ≥ 0.925). In hac comparative analysis, chi-quadratum test (McNerman test) ostendit quod in Sansure Biotech MMXIX-NCOV Assay results sunt significantly diversis a CRS results (P = 0.031) (Tabula II).
Ut patet in Fig.1 the percentage of lowest Ct value (< 20 Ct) of Abbott SARS-CoV-2 assay (combined RdRp and N gene) was 87.6% and ORF1a/b gene Ct value of Sansure Biotech 2019-nCoV assay showed that the percentage of low Ct value (< 20 Ct) was 50.3% and the high Ct value (36–40 Ct) was 3.2%. 1 the percentage of lowest Ct value (< 20 Ct) of Abbott SARS-CoV-2 assay (combined RdRp and N gene) was 87.6% and ORF1a/b gene Ct value of Sansure Biotech 2019-nCoV assay showed that the percentage of low Ct value (< 20 Ct) was 50.3% and the high Ct value (36–40 Ct) was 3.2%.Ut patet in Fig.I, процент наименьшего значения CT (<XX CT) анализа abbott Sars, Cov-II (комбинировавtил гRP и N) Составtил 87,6%, значение ct гена Orf1a / b анализа sansure biotech MMXIX, ncov показало что процент низзого зтоо процения CT (<XX ct) Сосокое зн6%, Высссс40 CT) Составляло 3,2%. 1, the percentage of the lowest Ct value (< 20 Ct) analysis of Abbott SARS-CoV-2 (combined gene RdRp and N) was 87.6%, and the Ct value of ORF1a/b gene analysis of Sansure Biotech 2019-nCoV showed that the percentage of low Ct value (< 20 Ct) accounted for 50.3%, and high value Ct (36–40 Ct) accounted Nam 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87.6%，Sansure Biotech 2019-nCoV 检测的ORF1a/b 基因Ct 值显示低Ct 值(< 20 Ct) 的百分比为50.3%，高Ct 值(36–40 Ct) 的百分比为3.2%。 As shown in Figure 1, the lowest Ct value percentage (< 20 Ct) of Abbott SARS-CoV-2 test (combination of RdRp and N gene) is 87.6%, the ORF1a/b gene Ct value of Sansure Biotech 2019-nCoV test shows low Ct值(< 20 Ct) 的 percentage is 50.3%, 高Ct 值(36–40 Ct) 的 percentage is 3.2%. Как показано на рисунке I, анализ abbott Sars, Cov-II (Сочетащий гены rdrp и N) имелентное Низкое ct (<XX CT) В размере 87,6%, а значение ct гена Orf1a / B в исследовании Sansure Biotech 2019- Нинкий NCOV показал ии показалалалий ct. Ut ostensum est in Figura I, in Abbott Sars, Cov-II Asay (combining RDRP et n genes) habebat in lowest percentage CT valorem (<XX CT - B Gene in Sansure Biotech MMXIX - In Analysis de NCoV ostendit humilis ct. Процент значений (<XX ct) Составил 50,3%, Аосокннт высоких Соцений ct (36-40 ct) Составил 3,2%. Et percentage of valores (<XX CT) erat 50.3%, et recipis summo ct values ​​(36-40 CT) erat 3.2%.Et Abbott Sars, Cov-II B Test memoriae CT values ​​super XXX. In alia manu, in BGI Sars-Cov-II Assay Orf1a / B Gene habebat princeps CT valorem (> XXXVI CT) percentage erat IV% (Fig. I). In alia manu, in BGI Sars-Cov-II Assay Orf1a / B Gene habebat princeps CT valorem (> XXXVI CT) percentage erat IV% (Fig. I). С другой стороны, в анализе bgi Sars, Cov-II ген orf1a / b имел высокое знавлчени CT (> XXXVI ct) процент которого Составлвля (IV которого составлвля (рис IV (рис. I). In alia manu, in analysis BGI Sars-Cov-II gene Orf1a / B habuit excelsum CT valorem (> XXXVI CT) et percentage of quem IV% (Fig. I).另一方面, 在 BGI Sars, Cov-II 检测中, Orf1a / B 基因具有高 ct 值 (> XXXVI CT) 的百分比为 IV% (图 I). In alia manu, in BGI Sars, Cov-II deprehendatur, in percentage of Orf1a / B Gene cum High CT Value (> XXXVI CT) is IV% (Figure I). С другой стороны, в Анализе bgi Sars, Cov-II процент Генов Orf1a / b с выкими значениями CT (IV% (составt IV (рис. I). In alia manu, in BGI Sars, Cov-II Analysis, in percentage of Orf1a / B gen High CT values ​​(> XXXVI CT) erat IV% (Fig. I).
In hoc studio, ut tulit CLXIV nasopharyngeal exempla. Nam omnia genera assays, RNA et amplificatio peragatur per modos et rhoncus suadetur ab respective manufacturers.
Hoc studium demonstrandum quod abbott scriptor test pro Sars, Cov-II habet eadem deprehensio perficientur ut CRS, cum C% positivum, negans et altiore concordantiam. Cohen scriptor Kappa consensu 1.00, significans plenum consensum cum CRS. A similis studio per University of Washington in US invenitur quod altiore sensibilitate et specifica de abbottio test pro Sars-COV-II erat XCIII% et C%, respectively, comparari ad Laboratory-determinari Assay (LDA) Cd. XI. Et Abbott Sars-Cov-II deprehensio ratio fundatur in simultaneous combined deprehendatur ad n et rdrp genes, ut tam genes magis sensitivo, obscuratis falsum negativas12. A studium in Viennam, Austria etiam ostendit quod magna extraction sixe volumina et deprehendatur eluentes voluminibus minimized MIXTURA effectus et auctus deprehendatur efficientia13. Ut, abbott 's perfecta par pro Sars-Cov-II assay potest consociata cum suggestus deprehensio ratio, ut simul deprehendat combinatoria genes, exstrahit numerus exemplar (0,5 ml), et utitur magna copia eluent (XL μl).
Nostra eventus etiam ostendit quod deprehensio perficientur de Daan geneticae test fuit fere idem quod de CRS. Hoc consistent cum studiis in Anhui University in HuaaNan, Sina et fabrica scriptor C% positivum consensu. Quamvis Renuntiationes consistent results, una sample erat falsum negative post retesting idem ELUICATUS, sed fuit positivum in abbott Sars-Cov, II et Sansure Biotech NCOV-MMXIX assays. Hoc insinuat quod sit variabilitatem in eventus trans diversis generibus assays. Nihilominus, in studio ferri ex in Sina15, effectus Daan Gene Asay erat significantly diversis (P <0.05) comparari ad Lab-defined Reference Assay. Nihilominus, in studio ferri ex in Sina15, effectus Daan Gene Asay erat significantly diversis (P <0.05) comparari ad Lab-defined Reference Assay. Тем не менее, в исследовании, п иоведенном в китае1151зза Daan gene знался анализно отличался (P <0,05) От их лабораторного эталонного Анализа. Sed in studio in Sina15, Daan gene scriptor analysis effectus fuit significantly diversis (P <0.05) ex officinarum reference analysis.然而, 在中国进行的研究中 XV, 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 XV, 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 генетического геста Daan значально отличались (P <0,05) по Сравнению С еГо эталонным лабораторным тестом. Tamen in studio in Sina15, eventus Daan scriptor geneticae test fuerunt significantly diversis (P <0.05) comparari ad suum Reference Laboratory test.Hoc discrepantia potest esse ex sensitivitate referat test ad deprehendere Sars, Cov- II, et ulteriores studiis sit amet determinare causam.
In addition, nostra studium aestimari comparativum perficientur de Sars, Cov-II BGI Assay cum CRS, showing praeclara positivum cento adipisci (PPA = 97.9%), negans percentage = (NPA = C% (Opa). ). = 98.8%). Cohen scriptor Kappa values ​​ostendit bona pactum (K = 0.975). Studies in Netherlands16 et Sinai ostensum est consistent results. Et Sars-Cov-II BGI test est unum gene (orf1a / b) deprehendatur test usura X μl amplificationem / deprehendatur. Quamvis bonum statistical cum nostris reference praecessi, in analysis desiderari duo positivum samples (1.22%) de totalis sample. Hoc potest habere ingens orci effectus in tradenda dynamics ad utrumque patientes estote et civitatem levels.
Alius Disputatio Analysis includitur in hoc studium erat in sansure biotech NCOV-MMXIX RRT-PCR (roo) asay; In altiore par percentage erat 96.3%. Et robore concordia fuit etiam determinari per Cohen Kappa valorem, quod erat 0.925, significans plenum consensum cum CRS. Iterum, nostri results sunt identical ad studiis deduxit ad Central South University in Changsha, Sina et ad orci Laboratory Department of Liuzhou Populi scriptor hospitalis, Liuzhou urbem, China17. Etiamsi in bonam statistical concordantiam memoriae, chi-quadratum test (Macnemar test) ostendit quod effectus in sansure Biotech asseus habuit in statistically differentia comparari CRS (P <0.005). Etiamsi in bonam statistical concordantiam memoriae, chi-quadratum test (Macnemar test) ostendit quod effectus in sansure Biotech asseus habuit in statistically differentia comparari CRS (P <0.005). Несмотря на то, что было зафиксировано уазанное Выланано Статискическое соответвие, критерий хи, критерий хи-критерий хи, критерий хи, критерий хи критератератй (критерий Макнемара) показал, что результат Анализа Sansure Biotech имеет Статистичиески знавнению с CRS по по савнению Сраличие (P < 0,005). Licet per bona statistical consensu supra est memoriae, chi-quadratum test (mcnemar test) ostendit quod effectus in sansure biotech habuit per legistically significant differentia comparari 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, 但 检验 ((Macnemar 检验 表明 ,, Sansure Biotech 检测 结果 与 CRS 相比 具有 显着 ((P <0.005 ...............))) 0.005 ...............))), 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 ...............))) 0.005 Несмотря на Отмеченное Выше хорошеееееетиствиеское Сответвие Соотвиеское Соотвие, критерий хи-квадрара) Показал Статистически значимую разницу (P <0,005) между анализом sansure biotech и Crs. Quamvis bonum statistical consensum attendendum supra, in Chi-quadratum test (McNamemar test) ostendit in statistically significant difference (P <0.005) inter Sansure Biotech et CRS.Sex samples (3.66%) inventa est falsum negativis comparari CRS (supplementum table I); Hoc ipsum, praesertim dynamica tradenda virum. Et supra notitia et sustinet hoc humilis deprehensio Rate15.
In hoc studio, ct values ​​sunt determinatae ad invicem asay et respective suggestus, cum infima media CT valore nuntiavit in abbott Sars, COV-II asay. Hoc eventum potest ad Abbotted est simultaneous combined geneticae temptationis ratio pro deprehendatur Sars-Cov- II. Ideo secundum Figura I, 87.6% of Abbott Sars, Cov-II results habuit CT values ​​infra XX. Tantum parva numerus of sample results (12.4%) in 20-30 range. CT valores super XXX non memoriae. In addition ut abbott usus est Sars-Cov-II panel geneticae testing format, hoc eventus potest esse ad inferioribus deprehendatur terminus (32.5 RNA exemplaria / ml. ml) XIX.
Hoc studium habet quidam limitations: Uno modo, non habent vexillum / reference modi [ut virales onus vel alius laboratorium probat (LDA)] ex defectu opibus. Secundo, omnes specimens in hoc studium erant Nasopharyngeal swabs, dum eventus non convenit aliis specimine genera, et tertio, nostralem magnitudine erat parvum.
Hoc studium comparari perficientur quattuor rart-PCR ad Sars-Cov- II per nasopharyngeal exempla. Omnes deprehendatur assisys prope comparabiles perficientur, cum exceptione in Sansure Biotech. Praeterea, humilis positivity rate identified in Sansure Biotech Assay ad CRS (P <0.05). Praeterea, humilis positivity rate identified in Sansure Biotech Assay ad CRS (P <0.05). Кроме того, В вылент низкий выявлен низкий процент положительных Сравнению с CRS (P <0,05). In addition, in Sansure Biotech test ostendit humilis percentage of positivum results comparari CRS (P <0.05).此外, 与 CS 相比, Sansure Biotech 检测的阳性率较低 (P <0.05).此外, 与 CS 相比, Sansure Biotech 检测的阳性率较低 (P <0.05). Кроме того, Анализ sansure biogroел более низкий уровень положительных результов по сравнению С CRS (P <0,05). In addition, in Sansure Biotech habuit inferiorem positivity rate comparari CRS (P <0.05).Et Sansure Biotech NCOV-MMXIX (Ruo) Analysis of PPA, NPA et altiore consensu exceditur 93.5% cum Cohen Kappa vires consensu valore 0.925. Denique, in Sansure Biotech (Ruo) necessitates adhuc validatio ad usum in Aethiopia, et additional investigationis debet considerari ad aestimare petat ex singulis manufacturers.
Comparativo studium consilio deduxit ad quattuor salutem facilities in addis Ababa, Eka Kotebe hospitalis, Millennium Ecclesia curatio Center, Zwooditus Memorial Hospitalis, et Sancti Petri Tuberculosis Vestibulum hospitalis. Et data est collecta inter Decembris I et XXXI, MMXX. Medical facilities hoc studio erant ex proposito electi secundum eorum altum numerum casibus et availability maioris curatio centra in urbe. Similiter instrumenta, inter (VII) D et abbott M2000 realis-vicis PCR instrumentis, electus secundum commendatione naat Reagent manufacturers, et quatuor PCR Dependences Kits sunt saltem quatuor. Gene test, abbott Sars, Cov-II test, sansure Biotech test et Sars, Cov-II BGI test per studium).
Testing pro Sars, Cov-II factum ex I ad XXX Decembris MMXXX usura III ml De Virisl Transport Medium (VTM) (Miraclean Technology, Shenzhen, Sinis) ab hominibus inquisitionis ad Covi-XIX Referred to Ephi. Nasopharyngeal exempla colligi exercitati sample collectores misit Ephi triplici sarcinas. Prior ad nucleic acidum solitudo, singulis specimen assignatur unicum idem numero. Extraction est ex singulis sample statim super adventu per manual et automatic extractionem modos. Sic, pro ipso extractionem abbott M2000, 1.3 ml (inter 0,8 ml Mortuus Volume et 0,5 ML extraction Inlet Volume) De Sample DNA ex singulis system (Abbott Molecular DNA et EXTRACTATIO SYMPART (Abbott Molecular Inc. DNA, USA). ) A batch de XCVI [XCII exempla, duo deprehendatur controls et duo non-template controls (NTC)] quod includitur in altiore processus (retrieval et deprehendatur) de duabus rounds of-COV-II (Eua) in Verus tempus. Mining. Similiter enim manual extraction, utere eodem exempla (nam automatic extraction et inventa). Ut per processum, CXL μl samples fuere aliquet et extrahendum usura Qiaamp viral RNA Mini Ornamentum (Qiagen GmbH, Hilden, Germania) in batches XXIV (including XX exemplar, duo rounds. Manually extractum eluat sunt amplificari et deprehendi per an abi (VII) D scelerisque cycler usura Sars-Cov-II BGI Assay, Daan Gene Asay, et Sansure Biotech.
Automated Solante et purificatio Sars-Cov-II viral RNA sequitur magnetica bead principle usura abbott DNA sample praeparatio reagentia. Inactivation of exempla et solubilization de virales particulas est ferri ex usura a purgat continet Guanidine Isothiscus ad denature dapibus et inactivate rnase. RNA est igitur separari a dapibus per solidum phase separationem per Silica, id est Guanidinium sal et alkaline PH de lysis buffer promovere binding of nucleic acida ad Silica (Sio2). Rinsing gradum remanet permanens proteins et obstantia ad producendum patet solutio. Dulcinum RNA est separatim ab Silica, secundum micryparticles per instrumentum scriptor magnetica Field20,21. In alia manu, manual segregatione et purificatio RNA est per se per nent columnam modum usura centrifugactione pro magnetica sto et separatio microparticas ex eluent.
Et Abbotted Real-Time Sars-Cov-II deprehendatur test (abbott Molecular, Inc.), quod factum est secundum fabrica instructiones, quae suscepit mala19,22 ab qui et FDA. In hac protocol, sample inactivation ante extractionem factum in aqua balneum ad LVI ° C ad XXX min. Post virum inactivation, nucleic acidum extraction factum est abbott M2000 sp instrumento a 0,5 ml VTM usura abbott M2000 DNA Sample Praeparatio. secundum fabrica. Amplificationem et deprehendatur peragantur usura abbott M2000 Rt-PCR instrumentum, et Dual deprehendatur quod factum est ad RDRP et n genes. Rox) et Vic P (proprietary mori) ad targeting et deprehendatur internum controls, permittens simultaneous deprehensio utrumque amplificationem products XIX.
Et amplificationem deprehendatur modum huius ornamentum fundatur in unum-step Rt-PCR technology. Et orf1a / B et n genes delecti ut conserventur regiones a Daan Gene technology ad deprehendere scopum regionem amplificationem. Imprimis primers et fluorescent probat (n gene probes intitulatum cum Fam, Orf1a / B Rimes intitulatum cum Vic), ut deprehendere Sars-Cov- II RNA in exemplaria. In ultima eluentes et Magister miscet parati per addendo V μl of eluent ad XX μl of Magistri misce ad ultima volumen XXV μl. Amplificationem et deprehendatur peragatur eodem in an abi (DCCL) XXXIV realis-vicis PCR instrumentum.
Et orf1a / B et n genes deprehenditur per Sansure Biotech NCOV-MMXIX nucleic acidum diagnostic Kit (fluorescent PCR Deprehensio). Para specifica probes pro se target gene eligendo et jam alma pro orf1a / B regionem et rox channel ad n gene. Ad hoc Asayum ornamentum, Eluent et Magistri misce reagentia sunt addidit ut sequitur: parare XXX μl Magistri mix Reagent et XX μl eluted sample pro deprehendatur / amplificationem. Verus-vicis PCR Abi (LXXV) XXV est propter amplificationem / deprehensio.
Et Sars-Cov-II BGI test est fluorescent realis-vicis RRT-PCR ornamentum ad diagnosis de Covi-XIX. In scopum regionem sita est in orf1a / B regione Sars-Cov- II, quod est unum gene deprehensio modum. Praeterea, humana domestica gene β-Actin est interiora regulatam target gene. Magistri misce est paratus a miscentes XX μl of Magistri misce reagent et X μl de extractum RNA Sample in bene Plate26. An abi (VII) D fluorescent quantitative realis-vicis PCR instrumentum esse propter amplificationem et deprehensio. Omnes nucleic acidum amplificationem, PCR currere condiciones ad se asay, et interpretatio eventus fuerunt secundum ad respective manufacturer scriptor instructiones (Tabula III).
In hac comparative analysis, ut non utor a referat vexillum modum determinare percent pactum (positivum, negans et altiore) et alia comparatione parametri ad quattuor analyzes. Quisque test comparationis factum est cum CRS, in hoc studium in CRS erat constitutus a regula 'aliquo positivum "et effectus determinari, non per unum test results. Praeterea, in casu de Covi-XIX Transmissus, falsum negative praecessi sunt periculo quam falsum positivum eventus. Ideo dicere 'positivum' quod accuratius quam possibile ex CRS eventum, saltem duo assaus probat esse positivum, significatione quod saltem unus positivum effectus est verisimile venire a Euas. Sic, ex quatuor test results, duo vel plures test eventus, qui date idem effectus consideretur vera positivum aut negative18,27.
Data erat collected usura exstructa notitia extraction formas, data ingressum et analysis peragantur usura Praecedo statistical software et SPSS version 23.0 enim descriptio statistics. Positivum, negans et altiore percent consensu sunt resolvitur, et kappa score est usus ad determinare gradum pactum cuiusque modum cum CRS. Kappa values ​​interpretantur ut sequitur: 0.01 ad 0.20 ad mitis pactum, 0,21 ad 0,40 ad generalem pactum, 0.41-0.60 et moderata pactum, 0.61-0.80 pro major pactum et 0.81-0.99 ad completum est in major pactum et 0.81-0.90 pro completa pacta.
Ethica Clearance est adeptus a University of Addis Ababa et omnes experimentalem protocols hoc studium erant probatus ab Aethiops Public Heel Salutem Institutum scriptor Scientific Ethic Review Board. Et Reference Number ad Ephi Ethic Licentia est Ephi / IRB, 279-2020. Omnes modi applicantur ad normam commendatione et praescripta Æthiops nationalibus comprehensive guidelines ad curatio de Covi-XIX. Insuper scripta conscio consensu adeptus ab omni studio participantium prior ad participationem in studiis.
Omnes notitia adeptus an resolvitur in hac studio sunt includitur in hoc editis articulum. Data supporting eventus huius studium sunt praesto ab respective auctor in rationabile petitionem.
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