Preliminary in silico assessment of the specificity of published molecular assays and design of new assays using the available whole genome sequences of 2019-nCoV

Updated to 1,467,158 high coverage COVID-19 WGS from GISAID using BLASTDB version 5.

Run Date: 06/15/2021

Table 1. PSET analysis using SARS-CoV-2 as the intended target. Perfect True Positive (PT), True Positive (TP), and False Negative (FN) counts for various assays were produced in each run. The five Noblis assays were compared alongside with assays produced in the US including the Corman (four assays) and CDC (three assays) brands. Assays originating from China, Hong Kong, Thailand, Japan, France, South Korea, and Singapore were also used in this comparison. Algorithms were set to penalize Ns on the subject sequence. Internal improvement to the PSET algorithms and snakemake (Snakemake — Snakemake 6.2.1 documentation 2) pipeline implementations decreased the Type I and Type II error rates compared with previous versions. We no longer calculate off-target hits due to the continued stability of these values.

Assay PT TP FN Percent True
Noblis.12 1428580 28171 261 99.98%
Noblis.40 1410187 46998 146 99.99%
Noblis.42 1442321 14775 54 100.00%
Noblis.44 1441381 15606 75 99.99%
Noblis.57 1436632 23166 172 99.99%
ncov_e_gene 1451168 5450 284 99.98%
ncov_n_gene 1416576 39005 300 99.98%
ncov_rdrp_1 0 1455502 1782 99.88%
ncov_rdrp_2 53 1456202 1029 99.93%
cdc_n1 1406276 50549 59 100.00%
cdc_n2 1406225 49444 180 99.99%
cdc_n3 1400673 56336 42 100.00%
China_N 385695 171479 898540 38.27%
China_ORF1ab 1445057 11579 224 99.98%
France_nCoV_IP2 1429891 27059 137 99.99%
France_nCoV_IP4 1350568 106392 327 99.98%
HKU-N 1419280 36071 303 99.98%
HKU-ORF1b-nsp14 1443698 13470 88 99.99%
Japan_NIID_2019-nCOV_N 3 1455163 327 99.98%
Japan_NIID_WH-1_F24381 1438097 21126 362 99.98%
Japan_NIID_WH-1_F501 1438212 18234 172 99.99%
Japan_NIID_WH-1_F509 1422997 33253 383 99.97%
Japan_NIID_WH-1_Seq_F24383 1437450 19715 2422 99.83%
Japan_NIID_WH-1_Seq_F519 1417852 25623 13148 99.10%
Japan_WuhanCoV-spk1 1444755 14483 329 99.98%
Thailand_WH-NIC_N 1431790 25005 104 99.99%
Chan-N 1413915 41254 247 99.98%
Chan-ORF1ab 52 1456204 272 99.99%
Chan-S 1402833 56135 1541 99.89%
Niu-E 1441802 14812 250 99.98%
Won-E 5 1456689 324 99.98%
Won-N 1418985 36687 125 99.99%
Won-ORF1ab 1414489 42714 27 100.00%
Won-S 1445481 11994 2752 99.81%
Yip-ORF1ab 1441625 14801 411 99.97%
Young-N 1 1456685 115 99.99%
Young-ORF1ab 1435631 21200 405 99.97%
Young-S 675594 58563 712969 50.73%
C1_COV_ORF1a 1425815 26184 254 99.98%
C2_COV_ORF1b 1418115 38997 73 99.99%
C3_COV_ORF3a 1305249 149194 703 99.95%
C4_COV_ORF8 716173 735316 2789 99.81%
C5_COV_S_gene 1437520 22364 193 99.99%