RT-PCR (Real time -PCR) in Coronavirus Disease 2019 (COVID-19) test
Diagnostic antibodies and antigens for Companion Animal disease testing
● Rabbit
Diagnostic antibodies and antigens for Swine disease testing
Diagnostic antibodies and antigens for Avian disease testing
Diagnostic antibodies and antigens for Multiple animal disease testing
Diagnostic antibodies and antigens for Ruminant disease testing
● Deer
Diagnostic antibodies and antigens for infectious and non-infectious Equine/Horse disease testing
SOCAIL MEDIA
Author:
Huajun Bai1 Xiaolong Cai1, 2* Xiaoyan Zhang1
1. R&D Center, GeneMedi Co.Ltd., Shanghai, P.R. China (www.genemedi.com)
2. Hanbio Research Center, Hanbio Tech Co. Ltd., Shanghai, P.R. China (www.hanbio.net)
Abstract:
The outbreak of COVID-19, caused by 2019 novel coronavirus (2019-nCoV), has been a global public health threat and caught the worldwide concern. Scientists throughout the world are sparing all efforts to explore strategies for the determination of the 2019-nCoV virus and diagnosis of COVID-19 rapidly. Several assays are developed for COVID-19 test , including RT-PCR, coronavirus antigens-based immunoassays, and CRISPR-based strategies (Cas13a or Cas12a), etc. Different assays have their advantages and drawbacks, and people should choose the most suitable assay according to their demands. Here, we make a brief introduction about these assays and give a simple overview of them, hoping to help doctors and researchers to select the most suitable assay for the Coronavirus Disease 2019 test (COVID-19 test) .
RT-PCR (Real time -PCR) in Coronavirus Disease 2019 (COVID-19) test
1) Principles for diagnostics
2) Advantages and disadvantages
Advantages:
① High sensitivity.
② Easy to be operated on a large scale.
Disadvantages:
① Professional technician and special apparatus are required to perform RT-PCR experiments and analyze the data, which costs so much.
② The standard positive control affects the experimental accuracy, and a false negative result may occur due to improper handling.
3) Optional Targets, primers, and probes from different departments
Different departments have their own systems for the determination of COVID-19 by targeting various open reading frames of SARS-CoV-2, which are shown as follows. Primers and probes of Orf1ab and N (nucleocapsid gene) are commonly used for COVID-19 test in China. Three pairs of primers and probes of N (nucleocapsid gene, N1, N2, N3) are applied in the US CDC for COVID-19. And primers and probes of E (envelope gene) are utilized for COVID-19 test in Europe.
| Organisation | Target | Forward primer (5’-3’) | Reverse primer (5’-3’) | Probe (5’-3’) |
University of Hong Kong, Beijing Center for Disease Prevention and Control, Capital Medical University Beijing Research Center for Preventive Medicine | orf1b | TGGGGYTTTACRGGTAACCT (Forward; Y=C/T, R=A/G) | AACRCGCTTAACAAAGCACTC (Reverse; R=A/G) | TAGTTGTGATGCWATCATGACTAG (Probe in 5’-FAM/ZEN/3’-IBFQ format; W=A/T) |
| N | TAATCAGACAAGGAACTGATTA | CGAAGGTGTGACTTCCATG | FAM-GCAAATTGTGCAATTTGCGG-IBFQ | |
Charité Virology, Berlin, Germany; Olfert Landt, Tib-Molbiol, Berlin, Germany; Erasmus MC, Rotterdam, The Netherlands; Public Health England, London | RdRP | GTGARATGGTCATGT GTGGCGG | CARATGTTAAASACA CTATTAGCATA | FAM-CAGGTGGA ACCTCATCAGGA GATGCBBQ |
| E gene | ACAGGTACGTTAATA GTTAATAGCGT | ATATTGCAGCAGTAC GCACACA | FAM-ACACTAGC CATCCTTACTGC GCTTCGBBQ | |
| N | CACATTGGCACCCGC AATC | GAGGAACGAGAAGA GGCTTG | FAM-ACTTCCTCA AGGAACAACATT GCCABBQ | |
The US Centers for Disease Control and Prevention (CDC), Integrated DNA Technologies | N1 | GACCCCAAAATCAGC GAAAT | TCTGGTTACTGCCAG TTG AATCTG | FAM-ACCCCGCA T TACGTTTGGTGG ACC-BHQ1 |
| N2 | TTACAAACATTGGCCGCAAA | GCGCGACATTCCGAAGAA | FAM-ACAATTTGCCCCCAGCGCTTCAG-BHQ1 | |
| N3 | GGGAGCCTTGAATACACCAAAA | TGTAGCACGATTGCAGCATTG | FAM-AYCACATTGGCACCCGCAATCCTG-BHQ1 | |
| RNAse P | AGATTTGGACCTGCGAGCG | GAGCGGCTGTCTCCACAAGT | FAM–TTCTGACCTGAAGGCTCTGCGCG–BHQ-1 |
Table 1. Optional Targets, primers, and probes from different departments
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