Evaluation of virological procedures to detect fetal human cytomegalovirus infection: Avidity of IgG antibodies, virus detection in amniotic fluid and maternal serum

Ruellan-Eugene, Genevieve; Barjot, Philippe; Campet, M.; Vabret, Astrid; Herlicoviez, M.; Muller, Georges; Levy, Gérard; Guillois, B.; Freymuth, F.

doi:10.1002/(sici)1096-9071(199609)50:1<9::aid-jmv3>3.0.co;2-5

Cited by 62 publications

(20 citation statements)

References 27 publications

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“…In fact, pp67 mRNA and IE mRNA NASBA assays showed 100 and 98.5% concordance, respectively, to DNA detection by PCR and virus isolation by the shell vial assay. In particular, all positive results obtained antenatally, with the exception of one IE mRNA-positive result, were confirmed at birth, thus supporting the high specificity of PCR results as previously reported by us and other groups (3,4,6,10,11,(14)(15)(16)(17). The discordant result obtained by the IE mRNA NASBA assay in the group of negative AF samples is difficult to explain.…”

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confidence: 83%

Prenatal Diagnosis of Congenital Human Cytomegalovirus Infection in Amniotic Fluid by Nucleic Acid Sequence-Based Amplification Assay

et al. 2003

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Nucleic acid sequence-based amplification assays for detection of human cytomegalovirus (HCMV) immediate-early and pp67 mRNA in 65 amniotic fluid samples tested for prenatal diagnosis of congenital HCMV infection showed sensitivity, specificity, and negative and positive predictive values >90%.Prenatal diagnosis of congenital human cytomegalovirus (HCMV) infection is mostly performed by rapid virus isolation from, and/or PCR detection of viral DNA in, amniotic fluid (AF) samples (1,3,11,12,14). In general, both techniques have been shown to provide high specificity (96 to 100%) and good sensitivity (70 to 90%) (4,8,10,14,15), with the exception of a few reports from a single group showing low specificity and positive predictive value for PCR on AF samples (5, 9). Since irrevocable decisions are often made on the basis of prenatal diagnosis, the use of a confirmatory assay is highly recommended. Thus, given the lack of cell culture facilities in many laboratories, molecular assays which may confirm PCR results for AF samples, thereby functioning as alternatives to HCMV isolation, must be identified.There are two commercially available assays for detection of HCMV immediate-early (IE) and late pp67 mRNA using the nucleic acid sequence-based amplification (NASBA) technique (BioMérieux, Boxtel, The Netherlands). The assay for pp67 mRNA is available as a complete kit (NucliSens CMV pp67), whereas a basic kit (NucliSens Basic Kit) is available for IE mRNA detection; IE mRNA-specific primer sequences can be obtained from BioMérieux. Currently, the NucliSens CMV pp67 assay is approved by the Food and Drug Administration for mRNA detection in blood, while the IE mRNA NASBA basic kit is to be used for research purposes.Overall, 65 AF samples from as many pregnant women with primary HCMV infection during pregnancy were tested. Primary infection was diagnosed by either HCMV-specific immunoglobulin G (IgG) seroconversion or the presence of HCMVspecific IgM associated with a low IgG avidity index (15). The virologic outcome of pregnancy was ascertained in newborns by HCMV isolation within the first 2 weeks of life and in aborted fetuses by virus isolation from and histologic examination of fetal tissues. All AF samples had been previously characterized for HCMV presence by both rapid virus isolation on shell vial cultures and viral DNA amplification by PCR (16).NASBA assays were performed retrospectively on AF samples according to the instructions given by the manufacturer for pp67 mRNA detection in blood samples (NucliSens CMV pp67). In particular, 49 AF samples collected from September 1991 through December 1999 and stored undiluted at Ϫ80°C were diluted 1:10 in NucliSens lysis buffer (BioMérieux) upon thawing. These samples were tested by NASBA for U1A mRNA, the transcript of a cellular gene with a low rate of transcriptional activity, to verify whether RNA was degraded during storage (7). The remaining 16 samples, collected after January 2000, were stored at Ϫ80°C as 1:10 dilutions in lysis buffer. Nucleic acids were extr...

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confidence: 83%

Prenatal Diagnosis of Congenital Human Cytomegalovirus Infection in Amniotic Fluid by Nucleic Acid Sequence-Based Amplification Assay

et al. 2003

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“…Avidity has been investigated among risk groups such as immunocompromised patients (40,(45)(46)(47), pregnant women (15,37,39,40), and neonates (48,49). Researchers have also studied avidity in immunocompetent persons (39,50).…”

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confidence: 99%

Cytomegalovirus Immunoglobulin G Avidity Index among Blood Donors in Alexandria, Egypt

Gawad

Hashish

Abaza

et al. 2016

Cent Eur J Public Health

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SUMMARYBackground and Aim: Transfusion transmitted diseases (TTD) are a major challenge to transfusion services all over the world. Cytomegalovirus (CMV) is considered one of the main viruses associated with blood transfusion. As CMV screening is not included in routine screening tests done for donated blood in blood banks in Egypt, the detection of CMV Immunoglobulin G (IgG) avidity needs to be tested for being a useful tool to diagnose recent infection among blood donors. The aim of this work was to study CMV IgG avidity index (AI) among blood donors.Methods: A total of 88 blood samples were collected from the non-remunerated volunteer blood donors who attended the Alexandria Regional Blood Transfusion Centre. A quantitative enzyme linked immunosorbent assay for the avidity detection of the specific IgG antibodies to CMV in human serum samples was used.Results: Eighty five studied blood donors (96.6%) were positive for CMV IgG. Eighty one donors (95.3%) showed high avidity (> 45.0%). Regarding the remaining four CMV IgG positive donors; three had medium avidity (< 45.0%) and only one had a low avidity of < 25.0%. A moderate agreement of 42.4% was found between IgG concentration and avidity.Conclusions: CMV seroprevalence was found to be high among volunteer blood donors, where age and gender were statistically significant factors associated with CMV IgG concentration. The use of the avidity assay as a screening tool for CMV among blood donors is highly suggested. The exclusion of the low and medium AI units will ensure the availability of a safe stock of blood units, hence eliminating the risk of CMV transmission to vulnerable groups.

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“…HCMV DNA detection in AF samples by qualitative PCR has been proposed as a useful tool for the prenatal diagnosis of HCMV congenital infection (4,14,18,33,36). However, this technique does not allow us to tell which infected fetuses will be symptomatic and which will remain asymptomatic.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, the direct detection of viral components, including pp65 antigenemia and DNAemia, can be helpful for acute infection diagnosis. Prenatal diagnosis of HCMV congenital infection relies on virus isolation from amniotic fluid (AF) and/or viral DNA detection by PCR in AF samples (4,12,24,25,36,42). In a previous study, we reported that the combination of culture and PCR on AF samples allows a reliable prenatal diagnosis, with 72% sensitivity and 97.6% specificity (18).…”

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confidence: 99%

Real-Time PCR Quantification of Human Cytomegalovirus DNA in Amniotic Fluid Samples from Mothers with Primary Infection

Gault²,

et al. 2002

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A real-time PCR assay was developed to quantify human cytomegalovirus (HCMV) DNA in amniotic fluid (AF) samples collected from 30 pregnant women with primary HCMV infection as detected either from HCMV-immunoglobulin G (IgG) seroconversion or by the presence of HCMV-specific IgG and IgM associated with a low IgG avidity. Clinical information available for each case included ultrasonographic examination and fetal or newborn outcome. HCMV infection of fetuses or newborns was confirmed for the 30 studied cases. AF samples were subdivided into three groups. In group A (n ‫؍‬ 13), fetuses presented major ultrasound abnormalities, and pregnancy was terminated. In group B (n ‫؍‬ 13), fetuses had normal ultrasound findings, the pregnancy went to term, and the newborns were asymptomatic at birth. In group C (n ‫؍‬ 4), fetuses had no or minor ultrasonographic signs, and pregnancy was terminated. The HCMV DNA load values in AF samples were significantly higher in group A (median, 2.8 ؋ 10 5 genome equivalents [GE]/ml) than in group B (median, 8 ؋ 10 3 GE/ml) (P ‫؍‬ 0.014). Our findings suggest that HCMV load level in AF samples correlates with fetal clinical outcome but might also be dependent on other factors, such as the gestational age at the time of AF sampling and the time elapsed since maternal infection.Human cytomegalovirus (HCMV) is the most common cause of viral intrauterine infection in developed countries, affecting 0.5 to 2% of all live births (1,32,40). Although the possibility of severe symptomatic fetal infection following recurrent maternal infection has been reported (8), fetal damage is mostly related to primary maternal infection (16,38,39). In this case, the transmission of the virus to the fetus may occur in 20 to 50% of pregnancies (18,27,38). It has been reported that HCMV transmission rates increase with gestational age and that a major risk of transmission is observed for seroconversion occurring late in pregnancy (5).Congenitally infected infants are asymptomatic at birth in about 90% of cases, and for symptomatic infants, infection ranges from mild to severe disseminated life-threatening disease resulting in up to 20% perinatal mortality (9,16,22,28,31,37). Up to 90% of the surviving symptomatic newborns will exhibit psychomotor and perceptual sequelae such as sensorineural hearing loss, mental retardation, cerebral palsy, seizures, and visual defects (2). Additionally, 10% of the infants who are asymptomatic at birth will later develop complications, mainly neurodevelopmental defects and deafness (7,15,40).The diagnosis of maternal primary HCMV infection is based mostly on serological testing, including HCMV immunoglobulin G (IgG) seroconversion and the presence of specific HCMV IgG and IgM (35). Moreover, the direct detection of viral components, including pp65 antigenemia and DNAemia, can be helpful for acute infection diagnosis. Prenatal diagnosis of HCMV congenital infection relies on virus isolation from amniotic fluid (AF) and/or viral DNA detection by PCR in AF samples (4,12,24,25,36,42)...

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Evaluation of virological procedures to detect fetal human cytomegalovirus infection: Avidity of IgG antibodies, virus detection in amniotic fluid and maternal serum

Cited by 62 publications

References 27 publications

Prenatal Diagnosis of Congenital Human Cytomegalovirus Infection in Amniotic Fluid by Nucleic Acid Sequence-Based Amplification Assay

Prenatal Diagnosis of Congenital Human Cytomegalovirus Infection in Amniotic Fluid by Nucleic Acid Sequence-Based Amplification Assay

Cytomegalovirus Immunoglobulin G Avidity Index among Blood Donors in Alexandria, Egypt

Real-Time PCR Quantification of Human Cytomegalovirus DNA in Amniotic Fluid Samples from Mothers with Primary Infection

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