One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2
Coronavirus disease 2019 (COVID‐19) has become pandemic since March 11, 2020. Thus, development and integration in clinics of fast and sensitive diagnostic tools are essential. The aim of the study is a development and evaluation of a one‐step quantitative reverse transcription‐polymerase chain reac...
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| Veröffentlicht in: | Journal of medical virology 2021-03, Vol.93 (3), p.1694-1701 |
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| container_title | Journal of medical virology |
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| creator | Goncharova, Ekaterina A. Dedkov, Vladimir G. Dolgova, Anna S. Kassirov, Ilia S. Safonova, Marina V. Voytsekhovskaya, Yana Totolian, Areg A. |
| description | Coronavirus disease 2019 (COVID‐19) has become pandemic since March 11, 2020. Thus, development and integration in clinics of fast and sensitive diagnostic tools are essential. The aim of the study is a development and evaluation of a one‐step quantitative reverse transcription‐polymerase chain reaction (RT‐qPCR) assay (COVID‐19 Amp) for severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) detection with an armored positive control and internal controls constructed from synthetic MS2‐phage‐based RNA particles. The COVID‐19 Amp assay limit of detection was 103 copies/ml, the analytical specificity was 100%. A total of 109 biological samples were examined using COVID‐19 Amp and World Health Organization (WHO)‐based assay. Discordance in nine samples was observed (negative by the WHO‐based assay) and discordant samples were retested as positive according to the results obtained from the Vector‐PCRrv‐2019‐nCoV‐RG assay. The developed COVID‐19 Amp assay has high sensitivity and specificity, includes virus particles‐based controls, provides the direct definition of the SARS‐CoV‐2 RdRp gene partial sequence, and is suitable for any hospital and laboratory equipped for RT‐qPCR.
Highlights
One‐step RT‐qPCR assay (COVID‐19 Amp) contains an armored positive control (ARC+) and an armored internal control (ICS) constructed from synthetic MS2‐phage‐based RNA particles.
Displays high specificity and selectivity rendering it a powerful diagnostic test
Suitable for any hospital and laboratory equipped for RT‐qPCR. |
| doi_str_mv | 10.1002/jmv.26540 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_7537076</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>2481909204</sourcerecordid><originalsourceid>FETCH-LOGICAL-c4710-4dbd734e4ce625929bd60e2e82095734370e00d33394ecadf3d285313fb63f153</originalsourceid><addsrcrecordid>eNp1kctuEzEUhi1ERUNhwQsgS2xgMe3xZTzxBimKuFWForR0a5zxGTrRzDi1Pamy4xH6jH0SDCkVILGxJZ_Pn_6jn5BnDA4ZAD9a9ZtDrkoJD8iEgVaFhoo9JBNgUhVKsXKfPI5xBQBTzfkjsi-4VkrJckK-ng54-_0mJlzTq9EOqU02tRuki_P8_Hm-oDZGu6XXbbqkNvQ-oKOLTzNa-yEF30Xa-EAdJqxT6wfqG3o2W5zlv3N_kU_-hOw1tov49O4-IF_evjmfvy9OTt99mM9OilpWDArplq4SEmWNipea66VTgBynHHSZB6ICBHBCCC2xtq4Rjk9LwUSzVKJhpTggr3fe9bjs0dWY49nOrEPb27A13rbm78nQXppvfmOqMrsrlQUv7wTBX40Yk-nbWGPX2QH9GA2XstSV1Fpn9MU_6MqPYcjrZWrKNGgOMlOvdlQdfIwBm_swDMzP3kzuzfzqLbPP_0x_T_4uKgNHO-C67XD7f5M5_nixU_4AJxekyQ</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2481909204</pqid></control><display><type>article</type><title>One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2</title><source>MEDLINE</source><source>Wiley Online Library Journals Frontfile Complete</source><creator>Goncharova, Ekaterina A. ; Dedkov, Vladimir G. ; Dolgova, Anna S. ; Kassirov, Ilia S. ; Safonova, Marina V. ; Voytsekhovskaya, Yana ; Totolian, Areg A.</creator><creatorcontrib>Goncharova, Ekaterina A. ; Dedkov, Vladimir G. ; Dolgova, Anna S. ; Kassirov, Ilia S. ; Safonova, Marina V. ; Voytsekhovskaya, Yana ; Totolian, Areg A.</creatorcontrib><description>Coronavirus disease 2019 (COVID‐19) has become pandemic since March 11, 2020. Thus, development and integration in clinics of fast and sensitive diagnostic tools are essential. The aim of the study is a development and evaluation of a one‐step quantitative reverse transcription‐polymerase chain reaction (RT‐qPCR) assay (COVID‐19 Amp) for severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) detection with an armored positive control and internal controls constructed from synthetic MS2‐phage‐based RNA particles. The COVID‐19 Amp assay limit of detection was 103 copies/ml, the analytical specificity was 100%. A total of 109 biological samples were examined using COVID‐19 Amp and World Health Organization (WHO)‐based assay. Discordance in nine samples was observed (negative by the WHO‐based assay) and discordant samples were retested as positive according to the results obtained from the Vector‐PCRrv‐2019‐nCoV‐RG assay. The developed COVID‐19 Amp assay has high sensitivity and specificity, includes virus particles‐based controls, provides the direct definition of the SARS‐CoV‐2 RdRp gene partial sequence, and is suitable for any hospital and laboratory equipped for RT‐qPCR.
Highlights
One‐step RT‐qPCR assay (COVID‐19 Amp) contains an armored positive control (ARC+) and an armored internal control (ICS) constructed from synthetic MS2‐phage‐based RNA particles.
Displays high specificity and selectivity rendering it a powerful diagnostic test
Suitable for any hospital and laboratory equipped for RT‐qPCR.</description><identifier>ISSN: 0146-6615</identifier><identifier>EISSN: 1096-9071</identifier><identifier>DOI: 10.1002/jmv.26540</identifier><identifier>PMID: 32966645</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>Adult ; Aged ; Aged, 80 and over ; Assaying ; Biological properties ; Biological samples ; Coronaviruses ; COVID-19 ; COVID-19 - diagnosis ; COVID-19 Testing - methods ; Diagnostic software ; Diagnostic systems ; Diagnostic Tests, Routine ; diagnostics ; Discordance ; Female ; Genome, Viral - genetics ; Humans ; Laboratories ; Male ; Middle Aged ; Molecular Diagnostic Techniques - methods ; Pandemics ; Phages ; Polymerase chain reaction ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Reverse transcription ; Ribonucleic acid ; RNA ; RNA phages ; RNA, Viral - genetics ; RNA-Dependent RNA Polymerase - genetics ; RT‐qPCR ; SARS-CoV-2 - genetics ; SARS-CoV-2 - isolation & purification ; SARS‐CoV‐2 ; Selectivity ; Sensitivity and Specificity ; Severe acute respiratory syndrome ; Severe acute respiratory syndrome coronavirus 2 ; Viral diseases ; Virology ; Viruses ; Young Adult</subject><ispartof>Journal of medical virology, 2021-03, Vol.93 (3), p.1694-1701</ispartof><rights>2020 Wiley Periodicals LLC</rights><rights>2020 Wiley Periodicals LLC.</rights><rights>2021 Wiley Periodicals LLC</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4710-4dbd734e4ce625929bd60e2e82095734370e00d33394ecadf3d285313fb63f153</citedby><cites>FETCH-LOGICAL-c4710-4dbd734e4ce625929bd60e2e82095734370e00d33394ecadf3d285313fb63f153</cites><orcidid>0000-0003-4736-2183 ; 0000-0002-4891-0439</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjmv.26540$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjmv.26540$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>230,314,776,780,881,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/32966645$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Goncharova, Ekaterina A.</creatorcontrib><creatorcontrib>Dedkov, Vladimir G.</creatorcontrib><creatorcontrib>Dolgova, Anna S.</creatorcontrib><creatorcontrib>Kassirov, Ilia S.</creatorcontrib><creatorcontrib>Safonova, Marina V.</creatorcontrib><creatorcontrib>Voytsekhovskaya, Yana</creatorcontrib><creatorcontrib>Totolian, Areg A.</creatorcontrib><title>One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2</title><title>Journal of medical virology</title><addtitle>J Med Virol</addtitle><description>Coronavirus disease 2019 (COVID‐19) has become pandemic since March 11, 2020. Thus, development and integration in clinics of fast and sensitive diagnostic tools are essential. The aim of the study is a development and evaluation of a one‐step quantitative reverse transcription‐polymerase chain reaction (RT‐qPCR) assay (COVID‐19 Amp) for severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) detection with an armored positive control and internal controls constructed from synthetic MS2‐phage‐based RNA particles. The COVID‐19 Amp assay limit of detection was 103 copies/ml, the analytical specificity was 100%. A total of 109 biological samples were examined using COVID‐19 Amp and World Health Organization (WHO)‐based assay. Discordance in nine samples was observed (negative by the WHO‐based assay) and discordant samples were retested as positive according to the results obtained from the Vector‐PCRrv‐2019‐nCoV‐RG assay. The developed COVID‐19 Amp assay has high sensitivity and specificity, includes virus particles‐based controls, provides the direct definition of the SARS‐CoV‐2 RdRp gene partial sequence, and is suitable for any hospital and laboratory equipped for RT‐qPCR.
Highlights
One‐step RT‐qPCR assay (COVID‐19 Amp) contains an armored positive control (ARC+) and an armored internal control (ICS) constructed from synthetic MS2‐phage‐based RNA particles.
Displays high specificity and selectivity rendering it a powerful diagnostic test
Suitable for any hospital and laboratory equipped for RT‐qPCR.</description><subject>Adult</subject><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>Assaying</subject><subject>Biological properties</subject><subject>Biological samples</subject><subject>Coronaviruses</subject><subject>COVID-19</subject><subject>COVID-19 - diagnosis</subject><subject>COVID-19 Testing - methods</subject><subject>Diagnostic software</subject><subject>Diagnostic systems</subject><subject>Diagnostic Tests, Routine</subject><subject>diagnostics</subject><subject>Discordance</subject><subject>Female</subject><subject>Genome, Viral - genetics</subject><subject>Humans</subject><subject>Laboratories</subject><subject>Male</subject><subject>Middle Aged</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Pandemics</subject><subject>Phages</subject><subject>Polymerase chain reaction</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>Reverse transcription</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>RNA phages</subject><subject>RNA, Viral - genetics</subject><subject>RNA-Dependent RNA Polymerase - genetics</subject><subject>RT‐qPCR</subject><subject>SARS-CoV-2 - genetics</subject><subject>SARS-CoV-2 - isolation & purification</subject><subject>SARS‐CoV‐2</subject><subject>Selectivity</subject><subject>Sensitivity and Specificity</subject><subject>Severe acute respiratory syndrome</subject><subject>Severe acute respiratory syndrome coronavirus 2</subject><subject>Viral diseases</subject><subject>Virology</subject><subject>Viruses</subject><subject>Young Adult</subject><issn>0146-6615</issn><issn>1096-9071</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kctuEzEUhi1ERUNhwQsgS2xgMe3xZTzxBimKuFWForR0a5zxGTrRzDi1Pamy4xH6jH0SDCkVILGxJZ_Pn_6jn5BnDA4ZAD9a9ZtDrkoJD8iEgVaFhoo9JBNgUhVKsXKfPI5xBQBTzfkjsi-4VkrJckK-ng54-_0mJlzTq9EOqU02tRuki_P8_Hm-oDZGu6XXbbqkNvQ-oKOLTzNa-yEF30Xa-EAdJqxT6wfqG3o2W5zlv3N_kU_-hOw1tov49O4-IF_evjmfvy9OTt99mM9OilpWDArplq4SEmWNipea66VTgBynHHSZB6ICBHBCCC2xtq4Rjk9LwUSzVKJhpTggr3fe9bjs0dWY49nOrEPb27A13rbm78nQXppvfmOqMrsrlQUv7wTBX40Yk-nbWGPX2QH9GA2XstSV1Fpn9MU_6MqPYcjrZWrKNGgOMlOvdlQdfIwBm_swDMzP3kzuzfzqLbPP_0x_T_4uKgNHO-C67XD7f5M5_nixU_4AJxekyQ</recordid><startdate>202103</startdate><enddate>202103</enddate><creator>Goncharova, Ekaterina A.</creator><creator>Dedkov, Vladimir G.</creator><creator>Dolgova, Anna S.</creator><creator>Kassirov, Ilia S.</creator><creator>Safonova, Marina V.</creator><creator>Voytsekhovskaya, Yana</creator><creator>Totolian, Areg A.</creator><general>Wiley Subscription Services, Inc</general><general>John Wiley and Sons Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7TK</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-4736-2183</orcidid><orcidid>https://orcid.org/0000-0002-4891-0439</orcidid></search><sort><creationdate>202103</creationdate><title>One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2</title><author>Goncharova, Ekaterina A. ; Dedkov, Vladimir G. ; Dolgova, Anna S. ; Kassirov, Ilia S. ; Safonova, Marina V. ; Voytsekhovskaya, Yana ; Totolian, Areg A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4710-4dbd734e4ce625929bd60e2e82095734370e00d33394ecadf3d285313fb63f153</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Adult</topic><topic>Aged</topic><topic>Aged, 80 and over</topic><topic>Assaying</topic><topic>Biological properties</topic><topic>Biological samples</topic><topic>Coronaviruses</topic><topic>COVID-19</topic><topic>COVID-19 - diagnosis</topic><topic>COVID-19 Testing - methods</topic><topic>Diagnostic software</topic><topic>Diagnostic systems</topic><topic>Diagnostic Tests, Routine</topic><topic>diagnostics</topic><topic>Discordance</topic><topic>Female</topic><topic>Genome, Viral - genetics</topic><topic>Humans</topic><topic>Laboratories</topic><topic>Male</topic><topic>Middle Aged</topic><topic>Molecular Diagnostic Techniques - methods</topic><topic>Pandemics</topic><topic>Phages</topic><topic>Polymerase chain reaction</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>Reverse transcription</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>RNA phages</topic><topic>RNA, Viral - genetics</topic><topic>RNA-Dependent RNA Polymerase - genetics</topic><topic>RT‐qPCR</topic><topic>SARS-CoV-2 - genetics</topic><topic>SARS-CoV-2 - isolation & purification</topic><topic>SARS‐CoV‐2</topic><topic>Selectivity</topic><topic>Sensitivity and Specificity</topic><topic>Severe acute respiratory syndrome</topic><topic>Severe acute respiratory syndrome coronavirus 2</topic><topic>Viral diseases</topic><topic>Virology</topic><topic>Viruses</topic><topic>Young Adult</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Goncharova, Ekaterina A.</creatorcontrib><creatorcontrib>Dedkov, Vladimir G.</creatorcontrib><creatorcontrib>Dolgova, Anna S.</creatorcontrib><creatorcontrib>Kassirov, Ilia S.</creatorcontrib><creatorcontrib>Safonova, Marina V.</creatorcontrib><creatorcontrib>Voytsekhovskaya, Yana</creatorcontrib><creatorcontrib>Totolian, Areg A.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Neurosciences Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of medical virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Goncharova, Ekaterina A.</au><au>Dedkov, Vladimir G.</au><au>Dolgova, Anna S.</au><au>Kassirov, Ilia S.</au><au>Safonova, Marina V.</au><au>Voytsekhovskaya, Yana</au><au>Totolian, Areg A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2</atitle><jtitle>Journal of medical virology</jtitle><addtitle>J Med Virol</addtitle><date>2021-03</date><risdate>2021</risdate><volume>93</volume><issue>3</issue><spage>1694</spage><epage>1701</epage><pages>1694-1701</pages><issn>0146-6615</issn><eissn>1096-9071</eissn><abstract>Coronavirus disease 2019 (COVID‐19) has become pandemic since March 11, 2020. Thus, development and integration in clinics of fast and sensitive diagnostic tools are essential. The aim of the study is a development and evaluation of a one‐step quantitative reverse transcription‐polymerase chain reaction (RT‐qPCR) assay (COVID‐19 Amp) for severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) detection with an armored positive control and internal controls constructed from synthetic MS2‐phage‐based RNA particles. The COVID‐19 Amp assay limit of detection was 103 copies/ml, the analytical specificity was 100%. A total of 109 biological samples were examined using COVID‐19 Amp and World Health Organization (WHO)‐based assay. Discordance in nine samples was observed (negative by the WHO‐based assay) and discordant samples were retested as positive according to the results obtained from the Vector‐PCRrv‐2019‐nCoV‐RG assay. The developed COVID‐19 Amp assay has high sensitivity and specificity, includes virus particles‐based controls, provides the direct definition of the SARS‐CoV‐2 RdRp gene partial sequence, and is suitable for any hospital and laboratory equipped for RT‐qPCR.
Highlights
One‐step RT‐qPCR assay (COVID‐19 Amp) contains an armored positive control (ARC+) and an armored internal control (ICS) constructed from synthetic MS2‐phage‐based RNA particles.
Displays high specificity and selectivity rendering it a powerful diagnostic test
Suitable for any hospital and laboratory equipped for RT‐qPCR.</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>32966645</pmid><doi>10.1002/jmv.26540</doi><tpages>0</tpages><orcidid>https://orcid.org/0000-0003-4736-2183</orcidid><orcidid>https://orcid.org/0000-0002-4891-0439</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of medical virology, 2021-03, Vol.93 (3), p.1694-1701 |
| issn | 0146-6615 1096-9071 |
| language | eng |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Adult Aged Aged, 80 and over Assaying Biological properties Biological samples Coronaviruses COVID-19 COVID-19 - diagnosis COVID-19 Testing - methods Diagnostic software Diagnostic systems Diagnostic Tests, Routine diagnostics Discordance Female Genome, Viral - genetics Humans Laboratories Male Middle Aged Molecular Diagnostic Techniques - methods Pandemics Phages Polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction - methods Reverse transcription Ribonucleic acid RNA RNA phages RNA, Viral - genetics RNA-Dependent RNA Polymerase - genetics RT‐qPCR SARS-CoV-2 - genetics SARS-CoV-2 - isolation & purification SARS‐CoV‐2 Selectivity Sensitivity and Specificity Severe acute respiratory syndrome Severe acute respiratory syndrome coronavirus 2 Viral diseases Virology Viruses Young Adult |
| title | One‐step quantitative RT‐PCR assay with armored RNA controls for detection of SARS‐CoV‐2 |
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