Top

Published in:

01-02-2006 | Original Article

The detection of CMV in amniotic fluid and cervicovaginal smear samples by real-time PCR assay in prenatal diagnosis

Authors: Aydan Biri, Æ Gulendam Bozdayı, Æ Banu C, Æ Bedia Dinc, Æ Aysegϋl Yucel, Æ Seyyal Rota

Published in: Archives of Gynecology and Obstetrics | Issue 5/2006

Abstract

Objective: There is no specific antiviral therapy or a vaccine, which could be safely administered to the pregnant women with primary human cytomegalovirus (CMV) infection. Therefore, prenatal diagnosis has a critical role in the management of pregnancy, complicated by this disease. In this study, we investigated the prevalence and clinical consequences of human CMV infection from cervicovaginal smear and amniotic fluid samples of pregnant women by using real-time polymerase chain reaction (RT-PCR) assay, in one of the Obstetrics and Gynecology outpatient clinics of Turkey. The identification of reliable prognostic markers of fetal disease remains the main purpose and a major challenge on this issue. Methods: Two hundred and six samples, of which 135 were cervicovaginal smear and 71 were amniotic fluid, were enrolled in the study. The DNAs of the samples were extracted by using Roche Diagnostic (Roche, Germany) kit and amplifications of these DNAs were studied by using Light-Cycler system (Roche Germany) as being quantitative. Anti-CMV IgM antibodies in the samples were studied by both MEIA (Imx system, Abbot Laboratories, USA) and a commercial ELISA kit (Radim SPA, Italy) while anti-CMV IgG antibodies were studied by MEIA (Axsym system, Abbot Laboratories, USA). Results: Human CMV DNA was found to be positive in 1.5% (2 in 135) of cervicovaginal smear and 1.4% (1 in 71) of amniotic fluid samples by RT-PCR. IgM and IgG were found to be negative in all of the cervicovaginal smear samples by both MEIA and ELISA, while IgG antibody was found to be positive in only one of the amniotic fluid samples by MEIA. Conclusion: With RT-PCR assay, we have found the prevalence of human CMV in pregnant women similar to epidemiologic reports, which have been described earlier. Whereas the fetus with positive amniotic fluid in favor of human CMV had an intrauterine growth restriction resulted in intrauterine exitus, no symptoms were observed in the infants of the other two pregnant women with positive RT-PCR results. The fact that the clinical consequence of the newborn whose amniotic fluid evaluation revealed human CMV infection by RT-PCR made us think that this molecular diagnosis method may be a reliable assay in prenatal diagnosis of this pathogen.

Andrews JI, Diekema DJ, Yankowitz J (2003) Prenatal testing for infectious disease. Clin Lab Med 23:295–315 Clinical manifestations, sonographic findings, diagnosis and treatment for clinically relevant congenital infections (such as CMV, parvovirus B19, varicella-zoster virus, Herpes simplex virus and T. gondii) are reviewed

Azam AZ, Vial Y, Fawer CL, Zufferey J, Hohlfeld P (2001) Prenatal diagnosis of congenital cytomegalovirus infection. Obstet Gynecol 97:443–445CrossRefPubMed

Barbi M, Binda S, Carappo V, Primache V, Dido P, Guidotti P (2001) CMV B genotypes and outcome of vertical transmission: study on dried blood spots of congenitally infected babies. J Clin Virol 21:75–79CrossRefPubMed

Bodeus M, Hubinont C, Bernard P, Bouckaer A, Thomas K, Goubau P (1999) Prenatal diagnosis of human cytomegalovirus by culture and polymerase chain reaction: 98 pregnancies leading to congenital infection. Prenat Diagn 19:314–317CrossRefPubMed

Bodeus M, Hubinont C, Goubau P (1999) Increased risk of cytomegalovirus transmission in utero during late gestation. Obstet Gynecol 93(5 Pt 1):658–660CrossRefPubMed

Boppana SB, Rivera LB, Fowler KB et al (2001) Intrauterine transmission of cytomegalovirus to infants of women with preconceptional immunity. N Engl J Med 344:1366–1371CrossRefPubMed

Catanzarite V, Dankner WM (1993) Prenatal diagnosis of congenital cytomegalovirus infection: false-negative amniocentesis at 20 weeks’ gestation. Prenat Diagn 13:1021–1025PubMedCrossRef

Chandler SH, Handsfield HH, McDougall JK (1987) Isolation of multiple strains of cytomegalovirus from women attending a clinic for sexually transmitted disease. J Infect Dis 155:655–660PubMed

Coonrod D, Collier AC, Ashley R, DeRouen T, Corey L (1998) Association between cytomegalovirus seroconversion and upper genital tract infection among women attending a sexually transmitted disease clinic: a prospective study. J Infect Dis 177:1188–1193PubMedCrossRef

10.

Demmler GJ, Buffone GJ, Schimbor CM, May RA (1988) Detection of cytomegalovirus in urine from newborns by using polymerase chain reaction DNA amplification. J Infect Dis 158:1177–1184PubMed

11.

Donner C, Liesnard C, Content J, Busine A, Aderca J, Rodesch F (1993) Prenatal diagnosis of 52 pregnancies at risk for congenital cytomegalovirus infection. Obstet Gynecol 82:481–486PubMedCrossRef

12.

Enders G, Bader U, Lindemann L, Schlasta G, Daiminger A (2001) Prenatal diagnosis of congenital cytomegalovirus infection in 189 pregnancies with known outcome. Prenat Diagn 21:362–377CrossRefPubMed

13.

Fisher S, Genbacev O, Maidji E, Pereira L (2000) Human Cytomegalovirus infection of plasental cytotrophoblasts in vitro and in utero: implications for transmission and pathogenesis. J Virol 74(15):6808–6820CrossRefPubMed

14.

Freij BJ, Sever JL (1999) Chronic infections. In: Avery GB, Fletcher MA, Mac Donald MG (eds) Neonatalogy: pathophysiology management of the newborn, 5th edn. Lippincott Williams-Wilkins, Philadelphia, pp 1123–1187

15.

Frukawa T, Jisaki F, Sakamuro D, Takegami T, Murayama T (1994) Detection of HCMV genome in uterus tissue. Arch Virol 135:265–277PubMedCrossRef

16.

Garcia AG, Fonseca EF, Marques RL, Lobato YY (1989) Placental morphology in cytomegalovirus infection. Placenta 10(1):1–18PubMedCrossRef

17.

Gouarin S, Gault E, Vabret A, Cointe D, Rozenberg F, Grangeot-Keros L et al (2002) Real-time PCR quantification of human cytomegalovirus DNA in amniotic fluid samples from mothers with primary infection. J Clin Microbiol 40(5):1767–1772PubMedCrossRef

18.

Griffiths PD, Stagno S, Pass RF, Smith RJ, Alford CA (1982) Congenital cytomegalovirus infection: diagnostic and prognostic significance of the detection of specific immunoglobulin M antibodies in cord serum. Pediatrics 69:544–549PubMed

19.

Griffiths PD, Baboonian C (1984) A prospective study of primary cytomegalovirus infection during pregnancy: final report. Br J Obstet Gynaecol 91(4):307–315PubMed

20.

Grose C, Weiner CP (1990) Prenatal diagnosis of congenital cytomegalovirus infection: two decades later. Am J Obstet Gynecol 163(2):447–450PubMed

21.

Guerra B, Lazzarotto T, Quarta S, Lanari M, Bovicelli L, Nicolosi A, Landini MP (2000) Prenatal diagnosis of symptomatic congenital cytomegalovirus infection. Am J Obstet Gynecol 183:476– 482PubMedCrossRef

22.

Hohlfeld P, Vial Y, Maillard-Brignon C (1991) Cytomegalovirus fetal infection; prenatal diagnosis. Obstet Gynecol 78:615–618PubMed

23.

Jones CA, Isaacs D (1995) Predicting the outcome of symptomatic congenital cytomegalovirus infection. J Pediatr Child Health 31:70–71

24.

Lazzarotto T, Guerra B, Spezzacatena P, Varani S, Gabrielli L, Pradelli P, Rumpianesi F, Banzi C, Bovicelli L, Landini MP (1998) Prenatal diagnosis of congenital cytomegalovirus infection. J Clin Microbiol 36:3540–3544PubMed

25.

Liesnard C, Donner C, Brancart F, Gosselin F, Delforge M-L, Rodesch F (2000) Prenatal diagnosis of congenital cytomegalovirus infection: prospective study of 237 pregnancies at risk. Obstet Gynecol 95:881–888CrossRefPubMed

26.

Lipitz S, Yagel S, Shalev E, Achiron R, Mashiach S, Schiff E (1999) Prenatal diagnosis of fetal primary cytomegalovirus infection. Obstet Gynecol 89:763–767CrossRef

27.

Mulongo KN, Lamy ME, Lierde MV (1995) Requirements for diagnosis of prenatal cytomegalovirus infection by amniotic fluid culture. Clin Diagn Virol 4:231–238CrossRefPubMed

28.

Nicolini U, Kustermann A, Tassis B, Fogliani R, Galimberti A, Percivalle E, Revello MG, Gerna G (1994) Prenatal diagnosis of congenital human cytomegalovirus infection. Prenat Diagn 14:903–906PubMedCrossRef

29.

Nigro G, Mazzocco M, Anceschi MM, La torre R, Antonelli G, Ermalando CV (1999) Prenatal diagnosis of fetal cytomegalovirus infection after primary or recurrent maternal infection. Obstet Gynecol 94:909–914CrossRefPubMed

30.

Revello MG, Baldanti F, Furione M, Sarasini A, Percivalle E, Zavzttoni M, Gerna G (1995) Polymerase chain reaction for prenatal diagnosis of congenital human cytomegalovirus infection. J Med Virol 47:462–466PubMedCrossRef

31.

Revello MG, Zavattoni M, Furione M, Baldanti F, Gerna G (1999) Quantification of HCMV DNA in amniotic fluid of mothers of congenitally infected fetuses. J Clin Microbiol 37:3350–3352PubMed

32.

Revello MG, Lilleri D, Zavattoni M et al (2003) Prenatal diagnosis of congenital human cytomegalovirus infection in amniotic fluid by nucleic acid sequence based amplification assay. J Clin Microbiol 41:1772–1774CrossRefPubMed

33.

Ruellan-Eugene B, Campet M, Varbet A, Herlicoviez M, Muller G, Levy G, Guillois B, Freymuth F (1996) Evaluation of virological procedures to detect fetal human cytomegalovirus infection: avidity of IgG antibodies, virus detection in amniotic fluid and maternal serum. J Med Virol 50:9–15CrossRefPubMed

34.

Sipewa MJ, Goubau P, Bodeus M (2002) Evaluation of a cytomegalovirus glycoprotein B recombinant enzyme immunoassay to discriminate between a recent and a past infection. J Clin Microbiol 40:3689–3693CrossRefPubMed

35.

Stagno S, Pass RF, Dworsky ME, Alford CA (1983) Congenital and perinatal cytomegalovirus infections. Semin Perinatol 7:31–42PubMed

36.

Stagno S, Whitley RJ (1985) Herpesvirus infections of pregnancy. Part I. Cytomegalovirus and Epstein-Barr virus infections. N Engl J Med 14:1270–1274CrossRef

37.

Stagno S (1995) Cytomegalovirus. In: Remington JS, Klein JO, editors. Infectious diseases of the fetus and newborn infant. Saunders, Philadelphia, pp 312–353

38.

Sohn YM, Oh MK, Balcarek KB, Cloud GA, Pass RF (1991) Cytomegalovirus infection in sexually active adolescents. J Infect Dis 163:460–463PubMed

39.

Yamamoto AY, Mussi-Pinhata MM, Pinto PCG, Figueiredo LTM, Jorge SM (2001) Usefulness of blood and urine samples collected on filter paper in detecting cytomegalovirus by the polymerase chain reaction technique. J Virol Methods 97:159–164CrossRefPubMed

40.

Zhou Y, Fisher SJ, Janatpour M, Genbacev O, Dejana E, Weelock M (1997) Human cytotrophoplasts adopt a vascular phenotype as they differentiate: a strategy for successful endovascular invasion? J Clin Invest 99:2139–2151PubMed

Title: The detection of CMV in amniotic fluid and cervicovaginal smear samples by real-time PCR assay in prenatal diagnosis
Authors: Aydan Biri
Æ Gulendam Bozdayı
Æ Banu C
Æ Bedia Dinc
Æ Aysegϋl Yucel
Æ Seyyal Rota
Publication date: 01-02-2006
Publisher: Springer-Verlag
Published in: Archives of Gynecology and Obstetrics / Issue 5/2006
Print ISSN: 0932-0067
Electronic ISSN: 1432-0711
DOI: https://doi.org/10.1007/s00404-005-0020-3

Keynote webinar | Spotlight on medication adherence

Springer Medicine

The detection of CMV in amniotic fluid and cervicovaginal smear samples by real-time PCR assay in prenatal diagnosis

Abstract

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 5/2006

Magnetic resonance imaging findings in leiomyoma of the ovary: a case report

Pseudo-Meig’s syndrome with multiple synchronous benign and malignant pelvic tumors

Epidermal cyst of the breast mimicking malignancy: clinical, radiological, and histological correlation

A study of abnormal cervical cytology in pregnant women

Diagnostic dilemmas in the delivery suite: myocardial ischemia due to coronary artery disease versus HELLP syndrome-related coronary artery spasm

Online manuscript submission for Archives of Gynecology and Obstetrics using the Editorial Manager system