tetano
Editor, Senior Moderator
Emerg Microbes Infect
. 2020 Jun 16;1-24.
doi: 10.1080/22221751.2020.1782774. Online ahead of print.
Development of an Automatic Integrated Gene Detection System for Novel Severe Acute Respiratory Syndrome-Related Coronavirus (SARS-CoV 2)
Yuchang Li[SUP] 1 [/SUP], Jing Li[SUP] 1 [/SUP], Ying Zhang[SUP] 1 [/SUP], Lizhong Dai[SUP] 2 [/SUP], Lin Li[SUP] 1 [/SUP], Juan Liu[SUP] 2 [/SUP], Sen Zhang[SUP] 1 [/SUP], Xiaoyan Wu[SUP] 1 [/SUP], Yi Hu[SUP] 1 [/SUP], Chenfeng Qin[SUP] 1 [/SUP], Tao Jiang[SUP] 1 [/SUP], Xiaoping Kang[SUP] 1 [/SUP]
Affiliations
Abstract
In December 2019, Wuhan, China suffered a serious outbreak of a novel coronavirus infectious disease (COVID) caused by novel severe acute respiratory syndrome-related coronavirus (SARS-CoV 2). To quickly identify the pathogen, we designed and screened primer sets, and established a sensitive and specific qRT-PCR assay for SARS-CoV 2; the lower limit of detection(LOD) was 14.8 (95% CI : 9.8-21) copies per reaction. We combined this qRT-PCR assay with an automatic integration system for nucleic acid extraction and amplification, thereby establishing an automatic integrated gene detection system (AIGS) for SARS-CoV 2. Cross reactive analysis performed in 20 other respiratory viruses and 37 nasopharyngeal swabs confirmed a 100% specificity of the assay. Using two fold diluted SARS-CoV 2 culture, the LOD of AIGS was confirmed to be 365copies/ml(95% CI: 351-375), which was Comparable to that of conventional q RT-PCR(740copies/ml, 95% CI: 689-750). Clinical performances of AIGS assay were assessed in 266 suspected COVID-19 clinical respiratory tract samples tested in parallel with a commercial kit. The clinical sensitivity of AIGS test was 97.62% (95%CI:0.9320-0.9951) based on the commercial kit test result , and concordance analysis showed a high agreement in SARS-CoV-2 detection between the two assays, Pearson R was 0.9623 (95% CI:0.9523-0.9703).The results indicated that this AIGS could be used for rapid detection of SARS-CoV 2. With the advantage of simple operation and less time consuming, AIGS could be suitable for SARS-CoV2 detection in primary medical institutions, thus would do a great help to improve detection efficiency and control the spread of COVID-19.
Keywords: SARS-CoV2; automatic integrated gene detection system; qRT-PCR.
. 2020 Jun 16;1-24.
doi: 10.1080/22221751.2020.1782774. Online ahead of print.
Development of an Automatic Integrated Gene Detection System for Novel Severe Acute Respiratory Syndrome-Related Coronavirus (SARS-CoV 2)
Yuchang Li[SUP] 1 [/SUP], Jing Li[SUP] 1 [/SUP], Ying Zhang[SUP] 1 [/SUP], Lizhong Dai[SUP] 2 [/SUP], Lin Li[SUP] 1 [/SUP], Juan Liu[SUP] 2 [/SUP], Sen Zhang[SUP] 1 [/SUP], Xiaoyan Wu[SUP] 1 [/SUP], Yi Hu[SUP] 1 [/SUP], Chenfeng Qin[SUP] 1 [/SUP], Tao Jiang[SUP] 1 [/SUP], Xiaoping Kang[SUP] 1 [/SUP]
Affiliations
- PMID: 32543298
- DOI: 10.1080/22221751.2020.1782774
Abstract
In December 2019, Wuhan, China suffered a serious outbreak of a novel coronavirus infectious disease (COVID) caused by novel severe acute respiratory syndrome-related coronavirus (SARS-CoV 2). To quickly identify the pathogen, we designed and screened primer sets, and established a sensitive and specific qRT-PCR assay for SARS-CoV 2; the lower limit of detection(LOD) was 14.8 (95% CI : 9.8-21) copies per reaction. We combined this qRT-PCR assay with an automatic integration system for nucleic acid extraction and amplification, thereby establishing an automatic integrated gene detection system (AIGS) for SARS-CoV 2. Cross reactive analysis performed in 20 other respiratory viruses and 37 nasopharyngeal swabs confirmed a 100% specificity of the assay. Using two fold diluted SARS-CoV 2 culture, the LOD of AIGS was confirmed to be 365copies/ml(95% CI: 351-375), which was Comparable to that of conventional q RT-PCR(740copies/ml, 95% CI: 689-750). Clinical performances of AIGS assay were assessed in 266 suspected COVID-19 clinical respiratory tract samples tested in parallel with a commercial kit. The clinical sensitivity of AIGS test was 97.62% (95%CI:0.9320-0.9951) based on the commercial kit test result , and concordance analysis showed a high agreement in SARS-CoV-2 detection between the two assays, Pearson R was 0.9623 (95% CI:0.9523-0.9703).The results indicated that this AIGS could be used for rapid detection of SARS-CoV 2. With the advantage of simple operation and less time consuming, AIGS could be suitable for SARS-CoV2 detection in primary medical institutions, thus would do a great help to improve detection efficiency and control the spread of COVID-19.
Keywords: SARS-CoV2; automatic integrated gene detection system; qRT-PCR.