Table 1.

Direct sequencing group—methods

Study (year)	Method	Number of specimens	Clone VE1 vendor	Dilution/incubation	Interpretation	Note
Bullock (2012)	Sanger	96	Spring Bioscience	NA/30 min	No intensity score. Positive if staining > 20% of tumor cells
Dvorak (2014)	Sanger	73	Ventana	NA/16 min	Intensity 0–3+; strong (3+), medium (2+), weak (1+) and no staining (0). Positive if unequivocal staining in > 85% tumor cells
Fisher (2013)	Pyrosequencing	25	Spring Bioscience	1:40/30 min	Intensity 0–3+; Positive if > 10% of tumor cells and 2+ or 3+. Anything less considered negative	Study also used pan-BRAF antibody. These were excluded from study
Ghossein (2013)	Mass spectrometry	31	Spring Bioscience	1:50/NA	Intensity 0–3+; 0 (no staining), 1+ (faint), 2+ (moderate), 3+ (strong) Considered positive if 2+
Kim (2014)	Sanger	91	Spring Bioscience	1:300/15 min	Allred
Kim (2014)	Pyrosequencing	91	Spring Bioscience	1:300/15 min	Allred
Kim (2018)	Sanger	697	Spring Bioscience	NA/32 min	No staining negative; weak/faint to strong/diffuse considered positive
Loo (2018)	Sanger	12	Ventana	NA/32 min	NA
Martinuzzi (2016)	Sanger	85	Spring Bioscience	1:50/NA	Capper et al. method
Oh (2018)	Sanger	23	Ventana	1:4/16 min	Diffuse homogenous staining in all tumor cells considered positive. Non-spec staining of colloid and equivocal weak or focal as negative
Qui (2015)	Sanger	127	Ventana	NA/16 min	NA
Zagzag (2013)	Sanger	37	Spring Bioscience	NA/NA	Intensity 0–3+ of 0–100% of tumor cells; < 1+ and < 20% was considered negative. > 80%, stratified into 1–3+ as positive	Some molecular results used to determine true positivity in weakly staining samples
Zhao (2019)	Sanger	185	Ventana	NA/16 min	Intensity 0–3; 1–3+ staining considered positive. No or only "slight" staining as negative
Zhu (2016)	Sanger	118	Spring Bioscience	NA/40 min	Intensity 0–3+; 0 if no or faint staining in 10% or fewer cells, 1+ faint > 10% cells, 2+ mod > 10% cells, 3+ strong > 10%. Positive when score 1+