International Urogynecology Journal

, Volume 25, Issue 9, pp 1237–1242 | Cite as

Collagen type III alpha 1 polymorphism (rs1800255, COL3A1 2209 G>A) assessed with high-resolution melting analysis is not associated with pelvic organ prolapse in the Dutch population

  • Sabrina L. LinceEmail author
  • Leon C. van Kempen
  • Jeroen R. Dijkstra
  • Joanna IntHout
  • Mark E. Vierhout
  • Kirsten B. Kluivers
Original Article


Introduction and hypothesis

The rs1800255, COL3A1 2209 G>A polymorphism in the alpha 1 chain of collagen type III has been associated with an increased risk of pelvic organ prolapse (POP). In one of our previous studies however, polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) misdiagnosed rs1800255, COL3A1 2209 G>A in 6 % of cases. The high-resolution melting (HRM) analysis on the contrary obtained a 100 % accordance for this specific polymorphism and was used in the present study to validate this risk factor for POP.


In this case–control study, women with and without symptoms of POP were included and compared. DNA was extracted from blood samples. HRM analysis was used to assess for the presence of the homozygous rs1800255. Groups were compared using the Pearson chi-square, Mann–Whitney, and t tests. The discrepancy between HRM and PCR-RFLP results was investigated using PCR-RFLP results available from our previous study.


The study included 354 women: 272 patients with POP and 82 controls; 18 (7 %) cases versus 3 (4 %) controls had a homozygous rs1800255, COL3A1 2209 G>A polymorphism (odds ratio 1.9, 95 % confidence interval 0.5–6.9, compared to the wild type), and thus no association between POP and the homozygous polymorphism could be demonstrated. A discrepancy between HRM and PCR-RFLP results was found in 8 % of the samples.


The previously found statistically significant association between the rs1800255, COL3A1 2209 G>A polymorphism as measured with PCR-RFLP and POP could no longer be demonstrated. This raises concerns regarding the results of other association studies using PCR-RFLP.


Polymorphism Single nucleotide Genetics Collagen type III High-resolution melting Pelvic organ prolapse Validation studies 


Conflicts of interest



  1. 1.
    Buchsbaum GM, Duecy EE, Kerr LA et al (2006) Pelvic organ prolapse in nulliparous women and their parous sisters. Obstet Gynecol 108(6):1388–1393PubMedGoogle Scholar
  2. 2.
    Buchsbaum GM, Duecy EE (2008) Incontinence and pelvic organ prolapse in parous/nulliparous pairs of identical twins. Neurourol Urodyn 27(6):496–498PubMedGoogle Scholar
  3. 3.
    Altman D, Forsman M, Falconer C et al (2008) Genetic influence on stress urinary incontinence and pelvic organ prolapse. Eur Urol 54(4):918–922PubMedGoogle Scholar
  4. 4.
    Nikolova G, Lee H, Berkovitz S et al (2007) Sequence variant in the laminin gamma1 (LAMC1) gene associated with familial pelvic organ prolapse. Hum Genet 120(6):847–856PubMedGoogle Scholar
  5. 5.
    Kluivers KB, Dijkstra JR, Hendriks JC et al (2009) COL3A1 2209 G>A is a predictor of pelvic organ prolapse. Int Urogynecol J Pelvic Floor Dysfunct 20(9):1113–1118PubMedGoogle Scholar
  6. 6.
    Chen HY, Chung YW, Lin WY et al (2008) Collagen type 3 alpha 1 polymorphism and risk of pelvic organ prolapse. Int J Gynaecol Obstet 103(1):55–58PubMedGoogle Scholar
  7. 7.
    Jeon MJ, Chung SM, Choi JR et al (2009) The relationship between COL3A1 exon 31 polymorphism and pelvic organ prolapse. J Urol 181(3):1213–1216PubMedGoogle Scholar
  8. 8.
    Martins KD, de Jármy-DiBella ZI, da Fonseca AM et al (2011) Evaluation of demographic, clinical characteristics, and genetic polymorphism as risk factors for pelvic organ prolapse in Brazilian women. Neurourol Urodyn 30(7):1325–1328Google Scholar
  9. 9.
    Chen HY, Lin WY, Chen YH et al (2010) Matrix metalloproteinase-9 polymorphism and risk of pelvic organ prolapse in Taiwanese women. Eur J Obstet Gynecol Reprod Biol 149(2):222–224PubMedGoogle Scholar
  10. 10.
    Chen C, Hill LD, Schubert CM et al (2010) Is laminin gamma-1 a candidate gene for advanced pelvic organ prolapse? Am J Obstet Gynecol 202(5):505.e1–505.e5Google Scholar
  11. 11.
    Chen HY, Chung YW, Lin WY et al (2008) Estrogen receptor alpha polymorphism is associated with pelvic organ prolapse risk. Int Urogynecol J Pelvic Floor Dysfunct 19(8):1159–1163PubMedGoogle Scholar
  12. 12.
    Chen HY, Wan L, Chung YW et al (2008) Estrogen receptor beta gene haplotype is associated with pelvic organ prolapse. Eur J Obstet Gynecol Reprod Biol 138(1):105–109PubMedGoogle Scholar
  13. 13.
    Chen HY, Chung YW, Lin WY et al (2009) Progesterone receptor polymorphism is associated with pelvic organ prolapse risk. Acta Obstet Gynecol Scand 88(7):835–838PubMedGoogle Scholar
  14. 14.
    Erali M, Voelkerding KV, Wittwer CT (2008) High resolution melting applications for clinical laboratory medicine. Exp Mol Pathol 85(1):50–58PubMedCentralPubMedGoogle Scholar
  15. 15.
    Pan M, Lin M, Yang L et al (2013) Glucose-6-phosphate dehydrogenase (G6PD) gene mutations detection by improved high-resolution DNA melting assay. Mol Biol Rep 40(4):3073–3082PubMedGoogle Scholar
  16. 16.
    Thomas V, Mazard B, Garcia C et al (2013) UGT1A1 (TA)n genotyping in sickle-cell disease: high resolution melting (HRM) curve analysis or direct sequencing, what is the best way? Clin Chim Acta 424:258–260PubMedGoogle Scholar
  17. 17.
    Ouragini H, Haddad F, Darragi I et al (2013) Rapid and inexpensive detection of common HBB gene mutations in Tunisian population by high-resolution melting analysis: implication for molecular diagnosis. Hematology 19(2):80–84PubMedGoogle Scholar
  18. 18.
    Marotta RV, Turri O, Morandi A et al (2011) High resolution melting analysis to genotype the most common variants in the HFE gene. Clin Chem Lab Med 49(9):1453–1457PubMedGoogle Scholar
  19. 19.
    Lince SL, Kluivers KB, Dijkstra JR et al (2009) Reliable identification of the type III collagen gene polymorphism rs1800255 with the use of high resolution melting analysis. Lab Med 40(10):604–606Google Scholar
  20. 20.
    Lince SL, van Kempen LC, Vierhout ME et al (2012) A systematic review of clinical studies on hereditary factors in pelvic organ prolapse. Int Urogynecol J 23(10):1327–1336Google Scholar
  21. 21.
    Peters DL, Barber RC, Flood EM et al (2003) Methodologic quality and genotyping reproducibility in studies of tumor necrosis factor −308G–>A single nucleotide polymorphism and bacterial sepsis: implications for studies of complex traits. Crit Care Med 31(6):1691–1696PubMedCentralPubMedGoogle Scholar
  22. 22.
    Jancik S, Drabek J, Berkovcova J et al (2012) A comparison of direct sequencing, pyrosequencing, high resolution melting analysis, TheraScreen DxS, and the K-ras StripAssay for detecting KRAS mutations in non small cell lung carcinomas. J Exp Clin Cancer Res 31:79PubMedCentralPubMedGoogle Scholar
  23. 23.
    Carbonell P, Turpin MC, Torres-Moreno D et al (2011) Comparison of allelic discrimination by dHPLC, HRM, and TaqMan in the detection of BRAF mutation V600E. J Mol Diagn 13(5):467–473PubMedCentralPubMedGoogle Scholar
  24. 24.
    Allen-Brady K, Cannon-Albright L, Farnham JM et al (2011) Identification of six loci associated with pelvic organ prolapse using genome-wide association analysis. Obstet Gynecol 118(6):1345–1353PubMedCentralPubMedGoogle Scholar
  25. 25.
    Lammers K, Lince SL, Spath MA et al (2012) Pelvic organ prolapse and collagen-associated disorders. Int Urogynecol J 23(3):313–319PubMedCentralPubMedGoogle Scholar

Copyright information

© The International Urogynecological Association 2014

Authors and Affiliations

  • Sabrina L. Lince
    • 1
    Email author
  • Leon C. van Kempen
    • 2
    • 3
  • Jeroen R. Dijkstra
    • 4
  • Joanna IntHout
    • 5
  • Mark E. Vierhout
    • 1
  • Kirsten B. Kluivers
    • 1
  1. 1.Department of Obstetrics and Gynecology (791)Radboud University Medical CenterNijmegenThe Netherlands
  2. 2.Department of PathologyMcGill UniversityMontrealCanada
  3. 3.Lady Davis Institute for Medical ResearchJewish General HospitalMontrealCanada
  4. 4.Department of PathologyRadboud University Medical CenterNijmegenThe Netherlands
  5. 5.Department for Health EvidenceRadboud University Medical CenterNijmegenThe Netherlands

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