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Table 5 Signatures predicted not to hit any target organisms

From: Predicting the sensitivity and specificity of published real-time PCR assays

 

Target Organism

Year

Analysis

[15]

Japanese encephalitis virus

2007

Too many mismatches in either forward or reverse primer. Several strains have 3 mismatches at 3' end of forward primer in addition to internal mismatches.

[15]

Yellow fever virus

2007

Reverse primer only has a blast hit to one strain (Angola71). Forward primer only has blast hits to 3 strains, and there are many mismatches (e.g. for Angola71, the 11 bases at the 3' end of the primer do not match).

[15]

Saint Louis encephalitis virus

2007

Too many mismatches in the reverse primer, with 3 mismatches at 3' end as well as at other locations.

[16]

Dengue virus 1

2006

Reverse primer does not have any BLAST hits to target.

[16]

Dengue virus 2

2006

Forward primer has 3 or 12 mismatches at 3' end for most strains, the probe has BLAST hits to only 7 of the 57 genomes available, and reverse primer only has a BLAST hit to 1 genome but there are 3 mismatches at the 3' end.

[16]

Dengue virus 4

2006

Too many mismatches in forward primer and in some cases the probe.

[10]

Dengue virus

2002

Too many mismatches in forward primer. However, they are at the 5' end, so assay could still work for some strains with 19 matches at the 3' end of the forward primer.

[12]

Pseudomonas aeruginosa

2004

No blast hits of probe to Pseudomonas aeruginosa

[12]

Bacteroides spp.

2004

Probe is not between or even in close proximity to the forward and reverse primers

[12]

Stenotrophomonas maltophilia

2004

No blast hits of probe to Stenotrophomonas maltophilia

[12]

Haemophilus influenzae

2004

Probe only matches in 17 of 22 bases, which is unlikely to give a strong signal, since probe is unlikely to bind prior to the primers as desired for real time TaqMan chemistry.