Multiplex testing for Factor II and Factor V mutations in thrombophilia: technical verification and clinical validation of the cobas® Factor II and Factor V test.

J Thromb Thrombolysis 2019 Jan;47(1):87-95

Roche Molecular Systems Inc, 4300 Hacienda Drive, Pleasanton, CA, 94588, USA.

Laboratory testing for thrombophilia is complicated but essential for diagnosis. In 2017, the cobas® Factor II and Factor V Test (cobas F2F5 test) was launched for use with the cobas z 480 analyzer. This qualitative polymerase chain reaction test enables multiplex Factor II and Factor V testing with flexible reporting and workflow efficiency. Here, we report the results from studies investigating the performance of the cobas F2F5 test. Technical performance verification, clinical validation, external laboratory performance, and workflow comparison studies were performed. Fresh and frozen whole-blood and genomic DNA (gDNA) samples were tested, and several manual and automated DNA isolation methods were used. Bidirectional Sanger sequencing was used to verify genotypes identified by the cobas F2F5 test. One hundred percent agreement between the cobas F2F5 test and Sanger sequencing was observed for all genotypes. An external laboratory using remnant clinical samples also yielded 100% agreement between cobas F2F5 test results and their routine testing method. The cobas F2F5 test reduced the total sample processing time compared with the LightCycler® 1.2 platform (98.6 vs 420.2 min; 96 samples). Hemoglobin, extraction buffer, and ethanol contamination of the gDNA sample can lead to invalid results. The cobas F2F5 test has a high degree of accuracy for identification of Factor II and Factor V genotypes. This multiplex testing with short sample processing time can reduce handling errors and increase efficiency. Both manual and automated DNA isolation methods can be used with the cobas F2F5 test.

Download full-text PDF

Source
http://link.springer.com/10.1007/s11239-018-1745-8
Publisher Site
http://dx.doi.org/10.1007/s11239-018-1745-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6336749PMC
January 2019
13 Reads

Publication Analysis

Top Keywords

cobas f2f5
32
f2f5 test
32
factor factor
20
test
11
factor
10
cobas
9
external laboratory
8
sanger sequencing
8
clinical validation
8
cobas® factor
8
dna isolation
8
isolation methods
8
factor test
8
verification clinical
8
f2f5
8
sample processing
8
processing time
8
automated dna
8
manual automated
8
agreement cobas
8

Similar Publications