Novel coronavirus and molecular diagnostic technique

doi:10.3877/cma.j.issn.2095-5820.2021.03.010

Abstract

Abstract:

Corona virus is a plus-single-stranded RNA virus, and currently there are 7 types of coronavirus known to cause diseases in human. Novel coronavirus (SARS-CoV-2) is a β genus coronavirus and mainly causes novel coronavirus pneumonia. At present, COVID-19 is rapidly spreading around the world and has become an emerging infectious diseases under regular prevention and control. In this paper, new techniques for genotyping and laboratory diagnosis of SARS-CoV-2, including real-time fluorescent quantitative polymerase chain reaction (FQ-PCR), loop-mediated isothermal amplification (LAMP), rapid diagnostic test of POCT immunochromatography, nucleic acid mass spectrometry (MALDI-TOF) and gene sequencing, were briefly reviewed.

Key words: Coronaviridae, Molecular classification, Gene

Lei Zhu, Qingyi Zhu. Novel coronavirus and molecular diagnostic technique[J]. Chinese Journal of Clinical Laboratory Management(Electronic Edition), 2021, 09(03): 174-181.

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URL: https://zhlcsysgldzzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-5820.2021.03.010

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References 38

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机构(Institute )	靶基因(Gene targets)
中国CDC(China CDC)	ORF 1ab和 N基因
法国巴斯德研究所(Pasteur，Paris，France)	2个RdRP靶基因，一个E基因
美国CDC(US CDC)	3个N靶基因
日本国立传染病研究所(NIID，Japan)	CoV多位点靶基因，S蛋白
德国柏林查理特大学(Charité，Germany)	RdRP，N基因
泰国国立卫生研究所(National Institute of Health，Thailand)	N基因

机构(Institute )	靶基因(Gene targets)
中国CDC(China CDC)	ORF 1ab和 N基因
法国巴斯德研究所(Pasteur，Paris，France)	2个RdRP靶基因，一个E基因
美国CDC(US CDC)	3个N靶基因
日本国立传染病研究所(NIID，Japan)	CoV多位点靶基因，S蛋白
德国柏林查理特大学(Charité，Germany)	RdRP，N基因
泰国国立卫生研究所(National Institute of Health，Thailand)	N基因

机构	靶基因	正向引物(5′-3′)	反向引物(5′-3′)	探针(5′-3′)
美国CDC	N基因	N1： GACCCCAAAATCAGCGAAAT N2： TTACAAACATTGGCCGCAAA N3： GGGAGCCTTGAATACACCAAAA RP-F RNAse： AGATTTGGACCTGCGAGCG	N1： TCTGGTTACTGCCAGTTGAATCTG N2： GCGCGACATTCCGAAGAA N3： TGTAGCACGATTGCAGCATTG RP-RRNAse： GAGCGGCTGTCTCCACAAGT	N1： FAM-ACCCCGCATTACGTTGTGGACC-BHQ1 N2： FAM-ACAATTEGCCCCCAGCGCTTCAG-BHQ1 N3： FAM-AYCACATTGGCACCCGCAATCCTG-BHO1 RP-P RNAse： FAM-TTCTGACCTGAAGGCTCTGCGCG-BHQ-1
中国CDC	ORF 1ab和N基因	ORF 1ab： CCCTGTGGGTTITACACTTAA N： GGGGAACITCTCCTGCTAGAAT	ORF 1ab： ACGATTGTGCATCAGCTGA N： CAGACATITTGCTCTCAAGCTG	ORF 1ab： FAM-CCGTCTGCGGTATGTGGAAAGGTTATGG-BHQ1 N： FAM-TITGCTGCTGCITGACAGATT-TAMRA
德国柏林查理特大学	RdRp.E.N基因	RGRp： GTGARATGGTCATGTGTGGCGG E： ACAGGTACGITTAATAGTTAATAGCGT	RdRp： CARATGTTAAASACACTATTAGCATA E： ATATEGCAGCAGTACGCACACA	RdRp l： FAM-CAGGTGGAACCTCATCAGGAGATGC-BBQ RdRp 2： FAM-CCAGGTGGWACRTCATCMMGGTGATGC-BBQ E： FAM-ACACTAGCCATCCETACTGCGCTTCG-BBQ
日本国立传染病研究所	N基因	N： AAATTTTGGGGACCAGGAAC	N： TGGCAGCTGTGTAGGTCAAC	N： FAM-ATGTCGCGCATTGGCATGGA-BHQ
泰国国立卫生研究所	N基因	N： CGTTTGGTGGACCCTCAGAT	N： CCCCACTGCGTTCTCCATT	N： FAM-CAACTGGCAGTAACCABOH1

机构	靶基因	正向引物(5′-3′)	反向引物(5′-3′)	探针(5′-3′)
美国CDC	N基因	N1： GACCCCAAAATCAGCGAAAT N2： TTACAAACATTGGCCGCAAA N3： GGGAGCCTTGAATACACCAAAA RP-F RNAse： AGATTTGGACCTGCGAGCG	N1： TCTGGTTACTGCCAGTTGAATCTG N2： GCGCGACATTCCGAAGAA N3： TGTAGCACGATTGCAGCATTG RP-RRNAse： GAGCGGCTGTCTCCACAAGT	N1： FAM-ACCCCGCATTACGTTGTGGACC-BHQ1 N2： FAM-ACAATTEGCCCCCAGCGCTTCAG-BHQ1 N3： FAM-AYCACATTGGCACCCGCAATCCTG-BHO1 RP-P RNAse： FAM-TTCTGACCTGAAGGCTCTGCGCG-BHQ-1
中国CDC	ORF 1ab和N基因	ORF 1ab： CCCTGTGGGTTITACACTTAA N： GGGGAACITCTCCTGCTAGAAT	ORF 1ab： ACGATTGTGCATCAGCTGA N： CAGACATITTGCTCTCAAGCTG	ORF 1ab： FAM-CCGTCTGCGGTATGTGGAAAGGTTATGG-BHQ1 N： FAM-TITGCTGCTGCITGACAGATT-TAMRA
德国柏林查理特大学	RdRp.E.N基因	RGRp： GTGARATGGTCATGTGTGGCGG E： ACAGGTACGITTAATAGTTAATAGCGT	RdRp： CARATGTTAAASACACTATTAGCATA E： ATATEGCAGCAGTACGCACACA	RdRp l： FAM-CAGGTGGAACCTCATCAGGAGATGC-BBQ RdRp 2： FAM-CCAGGTGGWACRTCATCMMGGTGATGC-BBQ E： FAM-ACACTAGCCATCCETACTGCGCTTCG-BBQ
日本国立传染病研究所	N基因	N： AAATTTTGGGGACCAGGAAC	N： TGGCAGCTGTGTAGGTCAAC	N： FAM-ATGTCGCGCATTGGCATGGA-BHQ
泰国国立卫生研究所	N基因	N： CGTTTGGTGGACCCTCAGAT	N： CCCCACTGCGTTCTCCATT	N： FAM-CAACTGGCAGTAACCABOH1

靶基因	引物名	引物序列5′-3′
ORF 1ab	正向引物F	5′-CCCTGTGGGTTTTAC ACTTAA-3′
	正向引物R	5′-ACGATTGTGCATCAGCTGA-3′
	荧光探针P	5′-FAM-CCGTCTGCGGTATGTGGAAAGGTTATGG-BHQ1-3′
N 基因	正向引物F	5′-GGGGAACTTCTCCTGCTAGAAT-3′
	正向引物R	5′-CAGACATTT TGCTCTCAAGCTG-3′
	荧光探针P	5′-FAM-TTGCTGCTGCTTGACAGATT-TAMRA-3′

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