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Moezzi L, Keshavarz Z, Ranjbaran R, et al. Fetal RHD Genotyping Using Real-Time Polymerase Chain Reaction Analysis of Cell-Free Fetal DNA in Pregnancy of RhD Negative Women in South of Iran. Int J Fertil Steril. 2016;10(1):62-70doi: 10.22074/ijfs.2016.4770.
Moezzi, L., Keshavarz, Z., Ranjbaran, R., Aboualizadeh, F., Behzad-Behbahani, A., Abdullahi, M., Ramezani, A., Samsami, A., & Sharifzadeh, S. (2016). Fetal RHD Genotyping Using Real-Time Polymerase Chain Reaction Analysis of Cell-Free Fetal DNA in Pregnancy of RhD Negative Women in South of Iran. International journal of fertility & sterility, 10(1), 62-70. https://doi.org/10.22074/ijfs.2016.4770
Moezzi, Leili, et al. "Fetal RHD Genotyping Using Real-Time Polymerase Chain Reaction Analysis of Cell-Free Fetal DNA in Pregnancy of RhD Negative Women in South of Iran." International journal of fertility & sterility vol. 10,1 (2016): 62-70. doi: https://doi.org/10.22074/ijfs.2016.4770
Moezzi L, Keshavarz Z, Ranjbaran R, Aboualizadeh F, Behzad-Behbahani A, Abdullahi M, Ramezani A, Samsami A, Sharifzadeh S. Fetal RHD Genotyping Using Real-Time Polymerase Chain Reaction Analysis of Cell-Free Fetal DNA in Pregnancy of RhD Negative Women in South of Iran. Int J Fertil Steril. 2016 Apr-Jun;10(1):62-70. doi: 10.22074/ijfs.2016.4770. Epub 2016 Apr 05. PMID: 27123202; PMCID: PMC4845531.
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Int J Fertil Steril. 2016 Apr-Jun;10(1):62-70. doi: 10.22074/ijfs.2016.4770. Epub 2016 Apr 05.

Fetal RHD Genotyping Using Real-Time Polymerase Chain Reaction Analysis of Cell-Free Fetal DNA in Pregnancy of RhD Negative Women in South of Iran.

International journal of fertility & sterility

Leili Moezzi, Zeinab Keshavarz, Reza Ranjbaran, Farzaneh Aboualizadeh, Abbas Behzad-Behbahani, Masooma Abdullahi, Amin Ramezani, Alamtaj Samsami, Sedigheh Sharifzadeh

Affiliations

  1. Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
  2. Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran; Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran.
  3. School of Advanced Medical Science and Technology, Shiraz University of Medical Sciences, Shiraz, Iran.
  4. Department of Obstetrics and Gynecology, Medical School, Shiraz University of Medical Sciences, Shiraz, Iran.

PMID: 27123202 PMCID: PMC4845531 DOI: 10.22074/ijfs.2016.4770

Abstract

BACKGROUND: Maternal-fetal RhD antigen incompatibility causes approximately 50% of clinically significant alloimmunization cases. The routine use of prophylactic anti-D immunoglobulin has dramatically reduced hemolytic disease of the fetus and newborn. Recently, fetal RHD genotyping in RhD negative pregnant women has been suggested for appropriate use of anti-D immunoglobulin antenatal prophylaxis and decrease unnecessary prenatal interventions.

MATERIALS AND METHODS: In this prospective cohort study, in order to develop a reliable and non-invasive method for fetal RHD genotyping, cell free fetal DNA (cffD- NA) was extracted from maternal plasma. Real-time quantitative polymerase chain reaction (qPCR) for detection of RHD exons 7, 5, 10 and intron 4 was performed and the results were compared to the serological results of cord blood cells as the gold standard method. SRY gene and hypermethylated Ras-association domain family member 1 (RASSF1A) gene were used to confirm the presence of fetal DNA in male and female fetuses, respectively.

RESULTS: Out of 48 fetuses between 8 and 32 weeks (wks) of gestational age (GA), we correctly diagnosed 45 cases (93.75%) of RHD positive fetuses and 2 cases (4.16%) of the RHD negative one. Exon 7 was amplified in one sample, while three other RHD gene sequences were not detected; the sample was classified as inconclusive, and the RhD serology result after birth showed that the fetus was RhD-negative.

CONCLUSION: Our results showed high accuracy of the qPCR method using cffDNA for fetal RHD genotyping and implicate on the efficiency of this technique to predict the competence of anti-D immunoglobulin administration.

Keywords: Cell-Free Fetal DNA; Prenatal Diagnosis; Real-Time Polymerase Chain Reaction

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