Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 4, pp 517–524 | Cite as

Genetic analysis of the M2/ANXA5 haplotype as recurrent pregnancy loss predisposition in the Malay population

  • Kai-Cheen Ang
  • Sushilnathan Kathirgamanathan
  • Ewe Seng Ch’ng
  • Yan-Yeow Lee
  • Anna-Liza Roslani
  • Bavanandan Naidu
  • Krishna Kumar
  • Ridzuan Abdullah
  • Siti-Nadiah Abdul Kadir
  • Narazah Mohd Yusoff
  • Wan Zaidah Abdullah
  • Nadja Bogdanova
  • Peter Wieacker
  • Arseni Markoff
  • Thean-Hock Tang



The aim of this study was to evaluate a new predisposition factor, M2/ANXA5 (RPRGL3), in recurrent pregnancy loss (RPL) patients of Malay origin, since it was previously known that the prevalence of this condition is relatively high among the Malay population of Malaysia, where conventional hereditary thrombophilia factors have been generally ruled out.


A total of 232 women who had experienced ≥2 unexplained RPL and 141 available male partners were recruited, with 360 healthy Malay and 166 parous female controls. Prevalence of M2 carriage and RPL odds ratios were calculated in (a) control and patient groups; (b) clinically defined subgroups in categories of pregnancy loss, primary, secondary, and tertiary; and (c) timing of pregnancy loss in early, ≤15th gestation week and “late” fetal losses, and >15th gestation week subgroups.


Both male and female subjects had similar M2/ANXA5 allele frequencies. The carrier rate of M2/ANXA5 for the general Malay population was 42.2 and 34.9% for parous controls. These carrier rates compared to Malay RPL subjects (52% M2 carriers) resulted in elevated odds ratios (95% confidence interval) of 1.53 (1.1 to 2.1) and 1.97 (1.3 to 3.1) accordingly for early fetal losses. Moreover, exceeding copy numbers of M2/ANXA5 alleles seemed to afflict a greater chance of RPL in couples, especially when both partners were M2 carriers.


This study confirmed the proposed role of M2/ANXA5 as embryonic, genetically associated thrombophilia predisposition factor for early RPL among ethnic Malay of Malaysia.


Annexin A5 M2/ANXA5 Recurrent pregnancy loss (RPL) Miscarriage 



The authors thank the women and their partners who participated in this study.

Compliance with ethical standards

The present genetic association study was approved by the Human Ethics Research Committee of the Universiti Sains Malaysia (USMKK/PPP/JEPeM [245.3.(2)]) and from the National Institutes of Health, Ministry of Health, Malaysia (NMRR-11-1044-9519). The study was carried out in accordance with The Code of Ethics of the World Health Organization (Declaration of Helsinki), and the criteria of strengthening the reporting of genetic association studies were observed as far as applicable. The volunteer subjects who agreed to participate have signed an informed consent before collection of peripheral blood samples. Random Malay population subjects were recruited at the Universiti Sains Malaysia, Penang campus from January 2011 to May 2013 with appropriate informed consent.


This work was supported by Universiti Sains Malaysia Research Universiti Grant (USM RU grant no: 1001/CIPPT/812100) awarded to TTH. AM was funded by a PI grant of the German Research Community, DFG, MA-6288/1-1. AKC was supported by MyBrain15 Program (KPM (b) 850304015158) under the Malaysian Ministry of Education.

Conflict of Interest

The authors declare that they have no conflict of interest.

Supplementary material

10815_2017_871_MOESM1_ESM.docx (55 kb)
Supplementary Table S1 (DOCX 54 kb)
10815_2017_871_MOESM2_ESM.docx (56 kb)
Supplementary Table S2 (DOCX 55 kb)
10815_2017_871_Fig3_ESM.gif (91 kb)
Supplementary Fig. S1

AS-PCR- lane 1: negative control (1st PCR); lanes 2 and 3: negative control (2nd PCR) for ‘normal’ and M2/ANXA5 specific primers; lanes 4: M2 heterozygous DNA template; lanes 5: DNA sample of a ‘normal’ genotype; lanes 6: M2 homozygous DNA template. Double lanes 4, 5 and 6: 1st track is specific for the ‘normal’ allele, 2nd track discriminates the M2 haplotype. (GIF 91 kb)

10815_2017_871_MOESM3_ESM.tiff (1.5 mb)
High resolution image (TIFF 1495 kb)


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Kai-Cheen Ang
    • 1
  • Sushilnathan Kathirgamanathan
    • 2
  • Ewe Seng Ch’ng
    • 1
  • Yan-Yeow Lee
    • 3
  • Anna-Liza Roslani
    • 4
  • Bavanandan Naidu
    • 5
  • Krishna Kumar
    • 3
  • Ridzuan Abdullah
    • 2
  • Siti-Nadiah Abdul Kadir
    • 6
  • Narazah Mohd Yusoff
    • 1
  • Wan Zaidah Abdullah
    • 7
  • Nadja Bogdanova
    • 8
  • Peter Wieacker
    • 8
  • Arseni Markoff
    • 8
  • Thean-Hock Tang
    • 1
    • 9
  1. 1.Advanced Medical and Dental InstituteUniversity Sains MalaysiaBertamMalaysia
  2. 2.Department of Obstetrics and GynaecologyHospital Sultan Abdul HalimSungai PetaniMalaysia
  3. 3.Department of Obstetrics and GynaecologyHospital Tuanku JaafarSerembanMalaysia
  4. 4.Department of Obstetrics and GynaecologyHospital Tengku Ampuan AfzanKuantanMalaysia
  5. 5.Department of Obstetrics and GynaecologyHospital Sultanah BahiyahAlor SetarMalaysia
  6. 6.Department of PathologyHospital Sultan Abdul HalimSungai PetaniMalaysia
  7. 7.Department of Haematology, School of Medical SciencesUniversiti Sains MalaysiaKubang KerianMalaysia
  8. 8.Institute of Human GeneticsUniversity of MuensterMuensterGermany
  9. 9.Advanced Medical and Dental Institute (AMDI)Universiti Sains MalaysiaKepala BatasMalaysia

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