Rapid Detection of Aneuploidies in Spontaneous Aborted Fetal Samples by Quantitative Fluorescence-PCR: A Descriptive Study

Document Type : Research Article

Authors

1 Department of Genetics, Sana Institute of Higher Education, Sari, Mazandaran, Iran

2 Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Mazandaran, Iran

Abstract

Chromosomal aneuploidies are the most chromosomal abnormalities at birth due to maternal meiosis I errors. Pregnancies with autosomal chromosomal aneuploidies that survive are namely trisomies 13 (Patau syndrome), 18 (Edward syndrome), and 21 (Down syndrome), account for 89% of chromosome abnormalities. Quantitative fluorescent polymerase chain reaction (QF-PCR) which amplifies specific DNA sequences called short tandem repeats (STRs), by using fluorescently labeled primers is a rapid technique for prenatal diagnosis of common aneuploidies. In this study, DNA extraction was performed from 100 samples isolated from muscle tissue of aborted fetuses. The analysis was performed by multiplex QF-PCR using a panel of 25 STRs markers for chromosomes X, Y, 13, 18, and 21. Our results showed that 20% of abortions were due to aneuploidy. 53% of mothers who had abortions were aged 26-35 years old and 32% of them were aged 36-45 years old. The analysis of muscle samples of aborted fetuses indicated that 20 samples showed chromosomal aneuploidy. Of the abnormal cases, 10 cases (~50 %) showed trisomy 21 followed by trisomy 18 (7 cases, ~35%), Klinefelter syndrome (2 cases, ~10 %), and showed trisomy X (1 case, ~5 %). Our results indicated that the D21S1414 marker showed the highest rate of heterozygosity in the study population. Besides some limitations of this study such as sample size, these results suggest that one of the causes of these abortions could be maternal age. We concluded that QF-PCR could be a rapid and reliable method to screen prenatal chromosomal aneuploidy and allow appropriate counseling.

Keywords

References

Ahangari N, Doosti M, Ahangari E, Baradarn Rafiee N, Ghayoor Karimiani E. 2016. Prenatal diagnosis of fetal aneuploidies using QF-PCR in 333 cases. Mol Med J 2(1): 38-42.
Aleyasin A, Ghazanfari M, Ganji SM, Jahanshad F. 2004. Application of molecular DNA markers (STRs) in molecular diagnosis of down syndrome in Iran. J Sci I R Iran15(2): 103-108.
Allen EG, Freeman SB, Druschel C, Hobbs CA, O'Leary LA, Romitti PA, Royle MH, Torfs CP, Sherman SL. 2009. Maternal age and risk for trisomy 21 assessed by the origin of chromosome nondisjunction: a report from the Atlanta and National Down Syndrome Projects. Hum Genet 125(1): 41-52.
Atef SH, Hafez SS, Mahmoud NH, Helmy SM. 2011. Prenatal diagnosis of fetal aneuploidies using QF-PCR: the egyptian study. J Prenat Med 5(4): 83-89.  
Badenas C, Rodríguez-Revenga L, Morales C, Mediano C, Plaja A, Pérez-Iribarne MM, Soler A, Clusellas N, Borrell A, Sánchez MA. 2010. Assessment of QF-PCR as the first approach in prenatal diagnosis. J Mol Diagn12(6): 828-834.
Bernatowicz K, Zimowski J, Łaczmańska I, Piotrowski K, Kashyap A, Bednarska-Makaruk M, Sąsiadek M, Gronwald J. 2019. Clinical utility of MLPA and QF-PCR techniques in the genetic testing of miscarriages. Russ J Genet 55(10):1259-1265.
Chitayat D, Langlois S, Wilson RD. 2007. Prenatal screening for fetal aneuploidy in singleton pregnancies. J Obstet Gynaecol Can33(7): 736-750.
Choi TY, Lee HM, Park WK, Jeong SY, Moon HS. 2014. Spontaneous abortion and recurrent miscarriage: a comparison of cytogenetic diagnosis in 250 cases. Obstet Gynecol Sci57 (6): 518-525.
Cirigliano V, Lewin P, Szpiro-Tapies S, Fuster C, Adinolfi M. 2001. Assessment of new markers for the rapid detection of aneuploidies by quantitative fluorescent PCR (QF-PCR). Ann Hum Genet 65(5): 421-427. Coelho, F, Marques, F, Gonçalves, M, Almeida VCO, Mateo ECC, Ferreira ACS. 2016. Detection of aneuploidies in spontaneous abortions by quantitative fluorescent PCR with short tandem repeat markers: a retrospective study. Genet Mol Res 15(3): gmr.15038617.
Comas C, Echevarria M, Carrera M, Serra B. 2010. Rapid aneuploidy testing versus traditional karyotyping in amniocentesis for certain referral indications. J Matern Fetal Neonatal Med 23(9): 949-955.
Diego-Alvarez D, Garcia-Hoyos M, Trujillo MJ, Gonzalez-Gonzalez, C, Rodriguez de Alba M, Ayuso C, Ramos-Corrales C, Lorda-Sanchez I. 2005. Application of quantitative fluorescent PCR with short tandem repeat markers to the study of aneuploidies in spontaneous miscarriages. Hum Reprod 20(5): 1235-1243.
Goud TM, Harassi SMA, Salmani KKA, Al Busaidy SM, Rajab A. 2009. Cytogenetic studies in couples with recurrent miscarriage in the Sultanate of Oman. Reprod BioMed Online 18(3): 424-429.
Gug C., Raţiu A, Navolan D, Drăgan I, Groza IM, Păpurică M, Vaida MA, Mozoş I, Jurcă MC. 2019. Incidence and spectrum of chromosome abnormalities in miscarriage samples: a retrospective study of 330 cases. Cytogenet Genome Res 158:171-183.
Hassold T, Abruzzo M, Adkins K, Griffin D, Merill M, Millie E, Saker D, Shen J, Zaragoza M. 1996. Human aneuploidy: incidence, origin, and etiology. Environ Mol Mutagen 28:167-175.
Hulten MA, Dhanjal S, Pertl B. 2003. Rapid and simple prenatal diagnosis of common chromosome disorders: advantages and disadvantages of the molecular methods FISH and QF–PCR. Reprod 126(3): 279-297.
Hunt P, Hassold T. 2010. Female meiosis: coming unglued with age. Curr Biol 20(17): R699-R702.
Khoshnood B, Pryde P, Wall S, Singh J, Mittendorf R, Lee KS. 2000. Ethnic differences in the impact of advanced maternal age on birth prevalence of down syndrome. Am J Public Health 90(11): 1778-1781.
Langlois S, Duncan A. 2011. Use of a DNA Method, QF-PCR, in the prenatal diagnosis of fetal aneuploidies. J Obstet Gynaecol Can 33(9): 955-960.
Lebedev IN, Ostroverkhova NV, Nikitina TV, Sukhanova NN, Nazarenko SA. 2003. Molecular cytogenetic characteristics of chromosome imbalance in spontaneous human abortion cells with low proliferative activity in vitro. Russ J Genet 39(8): 934-943.
Lee A, Kiessling AA. 2017. Early human embryos are naturally aneuploidy-can that be corrected? J Assist Reprod Genet34(1): 15-21.
Mann K, Donaghue C, Fox SP, Docherty Z, Mackie Ogilvie C. 2004. Strategies for the rapid prenatal diagnosis of chromosome aneuploidy. Eur J Hum Genet 12:907-915. Mann K, Ogilvie CM. 2012. QF-PCR: Application, overview and review of the literature. Prenat Diagn 32: 309-314.
Nagaoka SI, Hassold TJ, Hunt PA. 2012. Human aneuploidy: mechanisms and new insights into an age-old problem. Nat Rev Genet 13: 493-504.
Nicolini U, Lalatta F, Natacci F, Curcio C, Bui TH. 2004. The introduction of QF-PCR in prenatal diagnosis of fetal aneuploidies: time for reconsideration. Hum Reprod Update 10(6): 541-548.
Pal AK, Ambulkar PS, Waghmare JE, Wankhede V, Shende MR, Tarnekar AM. 2018. Chromosomal aberrations in couples with pregnancy loss: a retrospective study. J Hum Reprod Sci 11: 247-253.
Saadi A, Kushtgi P, Gopinath PM, Satyamoorthy K. 2010. Quantitative fluorescence polymerase chain reaction (QF-PCR) for prenatal diagnosis of chromosomal aneuploidies. Int J Hum Genet 10(1-3): 121-129.
Shaffer LG, Bui TH. 2007. Molecular cytogenetic and rapid aneuploidy detection methods in prenatal diagnosis. Am J Med Genet C Semin Med Genet145C: 87-98.
Shearer BM, Thorland EC, Carlson AW, Jalal SM, Ketterling RP. 2011. Reflex fluorescent in situ hybridization testing for unsuccessful product of conception cultures: a retrospective analysis of 5555 samples attempted by conventional cytogenetics and fluorescent in situ hybridization. Genet Med 13(6): 545-452.
Sherman SL, Freeman SB, Allen EG, Lamb NE. 2005. Risk factors for nondisjunction of trisomy 21. Cytogenet Genome Res 111:273-280.
Sloter E, Nath J, Eskenaz B, Wyrobek AJ. 2004. Effects of male age on the frequencies of germinal and heritable chromosomal abnormalities in humans and rodents. Fertil Steril81(4): 925-943.
Teles TMA, Paula CMMd, Ramos MG, Coxir SA, Penna MLF. 2017. Frequency of chromosomal abnormalities in products of conception. Rev Bras Ginecol Obstet 39 (3): 110-114.
van den Berg MMJ, van Maarle MC, van Wely M, Goddijn M. 2012. Genetics of early miscarriage. Biochim Biophys Acta 1822: 1951-1959.
Waters JJ, Waters KS. 1999. Trends in Cytogenetic prenatal diagnosis in the UK: results from UKNEQAS external audit, 1987-1998. Prenat Diagn (19): 1023-1026.
Wu Z, Liu N, Zhao Z, Kong X. 2016. Detection of chromosome aneuploidies in spontaneous abortion villus samples by quantitative fluorescence PCR. Chi J Med Genet 33(2):227-230.
Xu YW, Peng YT, Wang B, Zeng YH, Zhuang GL, Zhou CQ. 2011. High follicle-stimulating hormone increases aneuploidy in human oocytes matured in vitro. Fertil Steril95(1): 99-104.
Zhang XH, Qiu LQ, Ye YH, Xu J. 2017. Chromosomal abnormalities: subgroup analysis by maternal age and perinatal features in Zhejiang province of China, 2011-2015. Ital J Pediatr 43(1): 47.
Zhang T, Sun Y, Chen Z, Li T. 2018. Traditional and molecular chromosomal abnormality analysis of products of conception in spontaneous and recurrent miscarriage. BJOG 125(4):414-420.
Zou G, Zhang J, Li XW, He L, He G, Duan T. 2008. Quantitative fluorescent polymerase chain reaction to detect chromosomal anomalies in spontaneous abortion. Int J Gynaecol Obstet 103(3): 237-240.
Journal of Genetic Resources
Volume 7, Issue 2
September 2021
Pages 204-210

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