Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR
Bogdanov K.V.1 , Nikitin M.M.2, Slyadnev M.N.2
1. Institute of Hematology, V.A. Almazov Federal Medical Research Centre, St. Petersburg, Russia; Lumex Group, St. Petersburg, Russia 2. St. Petersburg State University, St. Petersburg, Russia; Lumex Group, St. Petersburg, Russia
Single nucleotide polymorphism (SNP) genotyping methods are widely used for the detection of hereditary thrombophilias caused by genetic defects in the coagulation system. The hereditary thrombophilias are frequently associated with higher incidences of point mutations in hemostasis (F2 20210G>A, F5 1691G>A) and folate metabolism (MTHFR 677C>Т, MTHFR 1298A>C) genes. Moreover, the combination of gene abnormalities in F2 or/and MTHFR with F5 Leiden mutation leads to increased risk of developing thrombosis. Thus, simultaneous detection of the multiple gene mutations in a sample has important clinical relevance. The microchip-based multiplex real time PCR for estimation of allele specific polymorphism in hemostatic and folate metabolism genes presented here has a high efficiency and may be used for laboratory diagnosis. The optimized protocol for estimation of 4 different types of genetic polymorphisms allowed PCR to be performed with minimal quantity of DNA template and PCR reagents including Taq polymerase and a short-term thermocycling.
Download PDF:
Keywords: hereditary thrombophilias, single nucleotide polymorphism, F2, F5, MTHFR, PCR
Citation:
Bogdanov K.V., Nikitin M.M., Slyadnev M.N. (2015) Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR. Biomeditsinskaya Khimiya, 61(3), 357-362.
Bogdanov K.V. et al. Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR // Biomeditsinskaya Khimiya. - 2015. - V. 61. -N 3. - P. 357-362.
Bogdanov K.V. et al., "Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR." Biomeditsinskaya Khimiya 61.3 (2015): 357-362.
Bogdanov, K. V., Nikitin, M. M., Slyadnev, M. N. (2015). Allele polymorphism analysis in coagulation factors F2, F5 and folate metabolism gene MTHFR by using microchip-based multiplex real time PCR. Biomeditsinskaya Khimiya, 61(3), 357-362.
Komarov A.L., SHakhmatova O.O., Rebrikov D.V., Trofimov D.YU, Kotkina T.I., Ilyushhenko T.A. (2011) Ratsional'naya Farmakoterapiya v Kardiologii, 7, 409-425. Scholar google search
Makatsariya A.D., Bitsadze V.O. (2003) Trombofilii i protivotromboticheskaya terapiya v akusherskoj praktike, Triada-KH, M., 496s. Scholar google search
Van de Water N.S., French J.K., Lund M., Hyde T.A., White H.D., Browett P.J. (2000) J. Am. Coll. Cardiol., 36, 717-722. CrossRef Scholar google search
Ridker P.M., Miletich J.P., Buring J.E., Ariyo A.A., Price D.T., Manson J.E., Hill J.A. (1998) Ann. Intern. Med., 128, 1000-1003. CrossRef Scholar google search
Kluijtmans L.A., van den Heuvel L.P., Boers G.H., Frosst P., Stevens E.M., van Oost B.A., den Heijer M., Trijbels F.J., Rozen R., Blom H.J. (1996) Am. J. Hum. Genet., 58, 35-41. Scholar google search
Grandone E., Margaglione M., Colaizzo D., D'Andrea G., Cappucci G., Brancaccio V., Di Minno G. (1998) Am. J. Obstet. Gynecol., 179, 1324-1328. Scholar google search
Lotta L.A., Wang M., Yu J., Passamonti S.M. (2012) BMC Med Genomics, 5, DOI:10.1186/1755-8794-5-7. CrossRef Scholar google search
Maggio A., Giambona A., Cai S.P., Wall J., Kan Y.W., Chehab F.F. (1993) Blood, 81, 239-242. Scholar google search
Wang J., Lin M., Crenshaw A., Hutchinson A., Hicks B., Yeager M. (2009) BMC Genomics, 10, DOI: 10.1186/1471-2164-10-561. CrossRef Scholar google search
Edwards K.J. (2004) in: Performing Real-Time PCR. In Real-Time PCR: An Essential Guide (Edwards K., Logan J., Saunders N., eds.), Horizon Bioscience, Norfolk, pp. 71-83. Scholar google search
Mansilha A., Araújo F., Severo M., Sampaio S.M., Toledo T., Albuquerque R. (2006) Phlebology, 21, 24-27. CrossRef Scholar google search
Gibson C.S., MacLennan A.H., Hague W.M., Haan E.A., Priest K., Chan A., Dekker G. A., Goldwater, P. N. (2005) Am. J. Obstetr. Gynecol., 193, 1437-1443. Scholar google search
Seipp M.T., Pattison D., Durtschi J.D., Jama M., Voelkerding K.V., Wittwer C.T. (2008) Clin. Chem., 54, 108-115. CrossRef Scholar google search
