Evaluation of the Nanosphere Verigene System and the Verigene F5/F2/MTHFR Nucleic Acid Tests.
BACKGROUND
Our ability to detect single nucleotide polymorphisms (SNPs) and gene mutations has become commonplace in the clinical laboratory setting. Molecular genetic testing for gene variants associated with hypercoagulability has become a standard of practice for Factor V and Factor II polymorphisms.


METHODS
In this study, we evaluated a novel technology that allows for the routine assessment of these SNPs, the Verigene System (Nanosphere Inc, Northbrook, IL), as a low-density array that does not require PCR amplification prior to detection. Precision was assessed by using multiple operators for within and between run performance evaluations. Accuracy was assessed by evaluating 176 DNA samples from patients who had been previously tested for the SNPs of interest in this multicenter study.


RESULTS
No mis-calls were made during the precision studies. Testing of the 176 DNA samples resulted in individual call rates for the F5, F2 and MTHFR genotypes of 98.3%, 94.9%, and 92.6%, respectively.


CONCLUSIONS
The Verigene F5/F2/MTHFR Nucleic Acid Tests for the Factor V (1691G>A), Factor II (20210G>A) and MTHFR (677C>T) genes were robust methods for SNP detection without the need for DNA amplification. The ease of use and performance of this system makes it suitable for the clinical laboratory setting.
Lefferts
Joel A
JA
Department of Pathology, Dartmouth Medical School, Dartmouth Hitchcock Medical Center and Norris Cotton Cancer Center, Lebanon, NH 03756, USA.
Jannetto
Paul
P
Tsongalis
Gregory J
GJ
D004247:DNA / Q000032:analysis* / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D005165:Factor V / Q000235:genetics*; D005838:Genotype; D006801:Humans; D042965:Methylenetetrahydrofolate Reductase (NADPH2) / Q000235:genetics*; D054118:Nanospheres; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D011516:Prothrombin / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005165:Factor V; D011516:Prothrombin; D004247:DNA; D042965:Methylenetetrahydrofolate Reductase (NADPH2)