VIASURE Rec Time PCR Detection x, eee eae ENGUSH __ 1, Intended use 3. Principle VASURE SaRs-co \VASURE SARS. CoV? Roa Tne PCR Detecton Kk detonad ore speci aent#icaten an cifereniaon of 2019 Novel Coronovin ISARS-CeV-2} fn expkoory samples om poten wih signs ond symptoms af CODA? infection. Th et onde for we 05 on odin tho lagnols of SARS-CoV-2 mn combination with crea and epidemiological ak factor. RNA & exacted fom raspioly specimens, ampifed wing R-PCR and coocieg ing Auerescent epee: eye probes spec for SARS-CO¥.2 VISURE SARS-Co ure ONA separating the < wich is propont CCoronevis are enveloped nonsegmented poitiveenie RNA vies and belong fo Coronado fom. there platonms 2. Summary and Explanation 0 sk cornowins speces tnown fo couse humon deaeias. Four vse: [227 OCES, NLD ond KK) couse commen cold symptoms ond the other hwo (iovers acute resokatory syndrome cornavive (SARS-COV) nd (Midcle Eost resckatery syndrome coranavius IMIERS-COV}) ore zoonotic on procucng mo severe ccomplcotont. SARE-COY ond MERS-CoV have couted more thon 10000 cumdsive cores nthe pat! wo decades wih mort tots of 3 MERS-CoV an 1K SARS-CoV. VASURE SARS Ce PCR asoy spec the palymeros ln December 2019, me people hal worked of of ved eround he Huanan sesloed mote! in Wuhan. Hubel etected in RC Province. China, have pietanted areumonia of unknown cows, Deep teavencing ania othe respKSIo eavioment somples indicated ¢ novel erenaving, which wor named fy 2019 novel cetonevius [201°C one liey SARS-CoV 4. Reage Humento-human tansmision of the SARS-CoV-2 hat Been confined even in the inubaion period witout | eo symptoms. ond the vit coueet severe rexptay less ke hose SARS-COV produced. Ahough the pneumonio| \ eer |s the principal ines sociated, « few patents have developed severe peumoria, pulroncry edema. 2c eee rosprotey astest ryndvome, or mutple organ fale ond death. Centar of Dkease Cont ond Prevention Sri CDC} beteves thot symptoms of SARS-Cov-2 may oppor In as few cs 2 doys oF a long os 14 days ctor ec cexpone, being the most common fever, cough. myagle and dyspnea. lets conven symtoms ar sre Woo pean headache, dlathea and vomiting. 1 seems thet people cbove 60 yeas OX, mals, ond people wih comorbistis most olen have severe dioose, iagnosis of SARS-CoV"? ls performed detectng conventional causes of pneumecia wat ond detected by rext- ‘genwrotion sequencing or eaHlime RT-PCR methods, Several ays tho! detec! he SARSCoV-2 have been are Curently avaiable, uch as China CDC \gene fargo, ORFlab and N), Charts ~ Germany (gone trae, RERP cond E) or US CDC Ithvee forgets in gene). WHO recommends upper respiotor oc! specimens (nosophorngeol and cropharyngeal swabs) ondor lower respiotoy specimens souls. endotracheal ospcte, or bronchodlveolrlovage in pokents wih more severe respitory diease) for he ideniicaton of SARSCo¥-2. In action. other cca specimens os blood une ond stool may be colected to moni Ihe presence of the vis }NCO212€ene50420 v.01 2VIASURE Real Time PCR Detection Kits 3. Principle of the procedure NASURE SARS-CoV-? Reo Tene PCR Delecton Kl designed forthe lagnots of SARS-CoV? in ‘espiatery caren aeston 8 dene ioe step real ine format where the evens tonscbton ana ne erence Gmeucoion of specif feget sequence occuin the sme action wel he ntted tts togera honaneoee senercing complementary ONA by revere rorsrptoxe which i folowed by tne amplfeaton oto conned ‘eden ct ORFIab and N genes for SARS-CoV-2 ung specie mes ond Ierecent ied wobe, VASURE SARS-COV.2 Real Tme PCR Detection Ki &bosed on the 5° exonucieaie activity of DNA polymerase Burn DNA ompiicaton, this ensme cleaves the probe bounded fo the comelemantoy DNA sequence. Seperating the quencher dye trom the teporer. Ths reaction generates on increase in the Auexexcent signal ‘whieh Is properfional fo the quantity of target template. Ths fuorescence can be mecsued on Real Teme PCR lattorns. VIASURE SARS-CoV.2 Real Time PCR Detection kit contoin in each wel all he components necessary for real ime PCR estoy (specific primes/probes. dNTPS. butter, polmerose and retroranserptase) in on stabilzed form. The ‘assay con use an Extraction Control {EC} which can be introduced into each sample at the iss butter stage of the extraction process. This contro! con be used fo moriter he extraction process andlor discard the inhibition of the polymerase activity. ORFlab gene is amplfed ond detected in FAM channel. N gene & omplfied and detected in ROX channel and the Extraction control [EC) in HEX chonnel. VIC or JOE channel [depending on the equipment used select the proper detection channel, ee Annex?) 4. Reagents provided VIASURE SARS-CoV-2 Real Time PCR Detection Ki includes the folowing materia ond reagents detoiled in Tables 1. 2 and 3. Based on the commercial presentation and the Real Time PCR platform used. the stabilaed PCR reaction mix could be placed inside aitferent well and could be marketed on mute formats, Table 1 includes material ond reagents to be used with 8-well stips compotibie devices [See Annex 1). Table 2 includes materials ond reagents to be used with 96.well plate compatible devices (See Annex 1). Table 3 includes materia’ ond reagents for use with Giagen/Cotbelt Rotor-Gene@ instruments for 4-wel sips.VIASURE Real Time PCR Detection Kits oe a haa Tescipton Testor | Amount “racer enzymes prrneprobes. | ovgtwtsips | _ bettas onvond ote ite | sn2xewelstio et siobiizos tomo Foarion to reconstuie The Renyarotion Blt rabies produc! tue || violx 8mt Tonfectious syrintic vophized {SARSCOV-2 Posive Contol Ne ed 1 viol Ton inectous nce oc ‘extraction Contol yoptitued Geen 1 wat Negative cont Ton jemplate contol Vow | wats tr Water RUASe/ONASE tee BNAse/ONASE ree Ware! write |_1violxt mt Sprcaicapa forseoing wel G70 [rsporent] 6/12x8-c00 Tearott cop sos peers rcnsporent| é/12x8-99.9%, respectively. oth results show high sensitivity and specifiy 1o detect SARS-CoV-2 ving VIASURE SARS-CoV? Real Te PCR 01 Detection Kit. 12.2. Analytical sensitivity Time PCR Det he tection mit of 210 RNA copies per reaction for ORF0B 1e PCR Detection Kit has « detection iit of 210 RNA copies Pr -action fo al Ti VIASURE SARS-COV-2 Re nd N genes (Figures 2. and 3). _arVIASURE Real Time PCR Detection Kits 1g C2184 Rech me PCR Detection Saver [FAM chennel nrvcses) Figere 3. Dovton series IN gene (107 uso) SE Scere nea Rens! 123. Analytical specificity The specificity of the SARS-Cov.2 assay was confirmed by testing @ panel consisting of cifferent microorganisms representing the most common respiratory pathogers, No crossteactvly wos detected between any of the folowing microorganisms tested.VIASURE Real Time PCR Detection Kils 123. Analytical specificity te specticiy ofthe SARS-COV? assy wos confab fesing Pane conte of diferent microorganisms vrererening the most common rexpratorypalnogens No crsvfenchly Wot detected belween any of the folowing microorganisms tested. T wanco2i2eenes0420 rev.) scenes TH spp= (v2 we X26 PONG HURRAY So A STET 12.4, Analytical reactivity The reactivity of VIASURE SARS-CoV-2 Real Te PCR Detection Ki was valle ogc NA em fumon 29 COV strain BetaCov/Germany/BavPetl/2020p.! showing postive ess ens ANNEX | compamiBlLiny W! fie 108 ow pr fe “en. ign pote sors ted‘a ern || Pm Speer} aa Bordetella bronchiseptica | Strepr oe eee ee aie | Chlamycia psittocr infuents A/New Coledoniaraan am genotypes A and C 2 virus. a Some | seamen || rmancacerge— rier tI (Wiens AI MCNSNTaSTaOTS aE Stetina ve | ORS NTT TO [ niverea a reraeree Tiueraa 8h Fomor Aenov SICA OTST sees ES {| sivwongen Human Bocavivs (2) vas (HINtjpcmop wns : naderanaar || os -aias | bers Ahoonomrcrtmearsme-—}——_Mssewar = | Seer eorrens | | RS shareware wescamens | Levene curt [| nensavetanioams naapy Sears a ee eae |= miamearoagamaT etfs | tegenete mcgaas! |) Bisena ea BS : Horan porcintuonea 1,2 Sond Legionelio pneumophia Vilvsraa A/Hong Kong/4@01/201@ WC : X26 (H3N2I ves SST | Moraxella catomhots Table & Reference pathogen Inlvenea A/ South Avshala/SS7ROTA WR 175 (HON) vis rontowsneee || 1 mictOOnganin ied ins say 12.4. Analytical reactivity The reactivity of VIASURE SAI Time P tumon 2019 evaluated ogcinst RNA fom Hi ime PCR Detection Kit was IRE SARS-CoV-2 Real Time }e reactivity of VIASUI COV strain BetaCoV/Germany/BavPatl /2020 p.1 showing postive resuts, st