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

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MOEZZI LEILI; KESHAVARZ ZEINAB; RANJBARAN REZA; ABOUALIZADEH FARZANEH; BEHZAD BEHBAHANI ABBAS; ABDULLAHI MASOOMA; RAMEZANI AMIN; SAMSAMI ALAMTAJ; SHARIFZADEH SEDIGHEH

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Journal: International Journal of Fertility & Sterility Year:2016 | Volume:10 | Issue:1 Page(s): 62-70

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Title

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

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Keywords

PRENATAL DIAGNOSIS

REAL-TIME POLYMERASE CHAIN REACTION

CELL-FREE FETAL DNA

Abstract

Background: Maternal-fetal RhD antigen incompatibility causes approximately 50% of clinically significant all oimmunization 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 (cffDNA) 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 hyper methylated 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.

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APA: Copy

MOEZZI, LEILI, KESHAVARZ, ZEINAB, RANJBARAN, REZA, ABOUALIZADEH, FARZANEH, BEHZAD BEHBAHANI, ABBAS, ABDULLAHI, MASOOMA, RAMEZANI, AMIN, SAMSAMI, ALAMTAJ, & SHARIFZADEH, SEDIGHEH. (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 AND STERILITY, 10(1), 62-70. SID. https://sid.ir/paper/305860/en

Vancouver: Copy

MOEZZI LEILI, KESHAVARZ ZEINAB, RANJBARAN REZA, ABOUALIZADEH FARZANEH, BEHZAD BEHBAHANI ABBAS, ABDULLAHI MASOOMA, RAMEZANI AMIN, SAMSAMI ALAMTAJ, SHARIFZADEH SEDIGHEH. 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 AND STERILITY[Internet]. 2016;10(1):62-70. Available from: https://sid.ir/paper/305860/en

IEEE: Copy

LEILI MOEZZI, ZEINAB KESHAVARZ, REZA RANJBARAN, FARZANEH ABOUALIZADEH, ABBAS BEHZAD BEHBAHANI, MASOOMA ABDULLAHI, AMIN RAMEZANI, ALAMTAJ SAMSAMI, and SEDIGHEH SHARIFZADEH, “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 AND STERILITY, vol. 10, no. 1, pp. 62–70, 2016, [Online]. Available: https://sid.ir/paper/305860/en

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