Evaluation of New Commercial Real-Time PCR Quantification Assay for Prenatal Diagnosis of Cytomegalovirus Congenital Infection

Abstract Human cytomegalovirus (HCMV) is the leading congenital infection agent in the world. The importance of screening this infection has been debated, as 10–15% of the asymptomatic newborns with HCMV at birth will present late sequelae. The aim of this study was to test the feasibility of using saliva pools from newborns in a screening program for congenital HCMV infection, in two Portuguese hospitals. The screening was based on the use of pools of 10 saliva samples for detection of viral DNA by real-time PCR. Whenever there was a positive pool, the samples were tested individually, and for each positive sample the result was confirmed with a urine sample collected in the first 2 weeks of life. The study involved 1492 newborns. One hundred and fifty pools were screened, with 14 positive results in saliva, but only 10 were confirmed in urine samples, giving a prevalence of congenital HCMV infection in both hospitals of 0.67% (CI95% 0.36 to 1.23%). Conclusion: The overall prevalence of congenital HCMV infection in both hospitals was 0.67%. The use of saliva pools proved to be effective for the screening of this congenital infection, allowing timely screening and confirmation in a large population, with associated cost reduction. What is Known:• Newborn screening for HCMV is desirable.• Saliva is a good and practical sample. What is New:• The feasibility of using saliva pools for a large-scale screening.• The cost reduction of this strategy.

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Real-time PCR versus shell vial culture on urine of patients with suspected congenital cytomegalovirus infection

Future Virology ◽

10.2217/fvl-2019-0078 ◽

2019 ◽

Vol 14 (9) ◽

pp. 585-591

Author(s):

Luana Coltella ◽

Stefania Ranno ◽

Giuseppe Pizzichemi ◽

Livia Piccioni ◽

Stefano Chiavelli ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Congenital Infection ◽

Urine Samples ◽

Congenital Cytomegalovirus ◽

Shell Vial ◽

Congenital Cmv Infection ◽

Polymerase Chain ◽

Improve Patient Management ◽

Improve Patient

Aims: Cytomegalovirus (CMV) is the most common cause of congenital infection. Aim of this study is to support quantitative real-time polymerase chain reaction (PCR) versus shell vials culture for CMV screening in urine samples. Patients & methods: A retrospective study was conducted on 255 urine samples belonging to patients admitted to Bambino Gesù Pediatric Hospital, Rome, Italy, with suspected congenital CMV infection. Results & conclusion: Quantitative real-time PCR resulted more standardized, faster, less operator-dependent, less laborious and most of all cost saving and more sensitive than shell vial culture. Since a negative result for CMV in urine means no congenital infection, a more sensitive tool for detection of CMV DNA is essential to improve patient management and to reduce healthcare costs associated to a late diagnosis.

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Simultaneous Detection of Trisomies 13, 18, and 21 with Multiplex Ligation-Dependent Probe Amplification–Based Real-Time PCR

Clinical Chemistry ◽

10.1373/clinchem.2010.146472 ◽

2010 ◽

Vol 56 (9) ◽

pp. 1451-1459 ◽

Cited By ~ 13

Author(s):

Qiwei Guo ◽

Yulin Zhou ◽

Xiaobo Wang ◽

Qingge Li

Keyword(s):

Prenatal Diagnosis ◽

Real Time ◽

Real Time Pcr ◽

Karyotype Analysis ◽

Simultaneous Detection ◽

Maternal Blood ◽

Trisomy 13 ◽

Clinical Samples ◽

Dependent Probe ◽

Single Reaction

BACKGROUND Trisomies 13, 18, and 21 account for the majority of chromosomal aneuploidies detected in prenatal diagnosis. Diagnosis of these trisomies relies mainly on karyotype analysis. Several molecular methods have been developed for trisomy detection, but performance or throughput limitations of these methods currently constrain their use in routine testing. METHODS We developed multiplex ligation-dependent probe amplification–based real-time PCR (MLPA/rtPCR) to simultaneously detect these 3 trisomy conditions with a single reaction. We applied the method to DNA isolated from 144 blinded clinical samples that included 32 cases of trisomy 21, 11 cases of trisomy 18, 1 case of trisomy 13, and 100 unaffected control samples; results were compared with karyotype analysis. RESULTS As judged by the results of the karyotype analysis, MLPA/rtPCR correctly detected all 44 cases of trisomy in the analysis of the blinded clinical samples. The method was able to detect a change in chromosome dosage as low as 1.2-fold. CONCLUSIONS This novel PCR-based technology simultaneously identified 3 types of trisomy in a single reaction and accurately detected trisomy with mosaicism, while reducing assay times and costs compared with conventional methods. The MLPA/rtPCR approach may have applicability in noninvasive prenatal diagnosis with maternal blood samples.

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Sensitivity of DCSR3/GAPDH Ratio Using Quantitative Real-Time PCR in the Rapid Prenatal Diagnosis for Down Syndrome

Fetal Diagnosis and Therapy ◽

10.1159/000218031 ◽

2009 ◽

Vol 25 (2) ◽

pp. 220-223 ◽

Cited By ~ 5

Author(s):

Sanaa M.H. Helmy ◽

Somaya Ismail ◽

Randa Bassiouni ◽

Khaled R. Gaber

Keyword(s):

Down Syndrome ◽

Prenatal Diagnosis ◽

Real Time ◽

Real Time Pcr ◽

Quantitative Real Time Pcr

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Novel real-time PCR assay for rapid prenatal diagnosis of Down syndrome: A prospective study of 563 amniocytes

Clinical Biochemistry ◽

10.1016/j.clinbiochem.2009.02.004 ◽

2009 ◽

Vol 42 (7-8) ◽

pp. 672-675 ◽

Cited By ~ 3

Author(s):

Yu-ning Zhu ◽

Shi-ming Lu ◽

Jian-fei You ◽

Bo Zhu ◽

Mei-yu Yu

Keyword(s):

Down Syndrome ◽

Prenatal Diagnosis ◽

Prospective Study ◽

Real Time ◽

Real Time Pcr ◽

Pcr Assay ◽

A Prospective Study

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A One-Step Real-Time PCR Assay for Rapid Prenatal Diagnosis of Sickle Cell Disease and Detection of Maternal Contamination

Molecular Diagnosis ◽

10.1007/bf03260020 ◽

2003 ◽

Vol 7 (1) ◽

pp. 45-48 ◽

Cited By ~ 5

Author(s):

Catherine Costa ◽

Serge Pissard ◽

Emmanuelle Girodon ◽

Danièle Huot ◽

Michel Goossens

Keyword(s):

Prenatal Diagnosis ◽

Sickle Cell Disease ◽

Real Time ◽

Sickle Cell ◽

Real Time Pcr ◽

Cell Disease ◽

Pcr Assay ◽

One Step ◽

Maternal Contamination

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

Archives of Gynecology and Obstetrics ◽

10.1007/s00404-005-0020-3 ◽

2005 ◽

Vol 273 (5) ◽

pp. 261-266 ◽

Cited By ~ 8

Author(s):

Aydan Biri ◽

Æ Gulendam Bozdayı ◽

Æ Banu C ◽

Æ Bedia Dinc ◽

Æ Aysegϋl Yucel ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Amniotic Fluid ◽

Real Time ◽

Real Time Pcr ◽

Pcr Assay

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Fast-track strategy for the prevention of Hb Bart’s hydrops fetalis syndrome

Thalassemia Reports ◽

10.4081/thal.2017.6620 ◽

2017 ◽

Vol 7 (1) ◽

Author(s):

Narutchala Suwannakhon ◽

Khajohnsilp Pongsawatkul ◽

Teerapat Seeratanachot ◽

Wirawan Rasri ◽

Khwanruedee Mahingsa ◽

...

Keyword(s):

At Risk ◽

Prenatal Diagnosis ◽

Real Time ◽

Real Time Pcr ◽

Fast Track ◽

Dna Analysis ◽

Hydrops Fetalis ◽

Quantitative Real Time Pcr ◽

Risk Detection ◽

Early Risk

We propose a fast-track strategy [direct blood DNA analysis using a quantitative real-time polymerase chain reaction (PCR) technique] for the early risk detection and prenatal diagnosis of α(0)-thalassemia (SEA and Thai deletion). Blood DNA samples were obtained from a volunteer group of 1235 ANC couples. They were assessed using quantitative real-time PCR to detect carriers of α(0)-thalassemia (SEA and Thai deletion). At-risk couples were identified, and further prenatal diagnosis by amniocentesis was implemented. Fetal DNA was isolated from the amniotic cells and characterized by quantitative real-time PCR to detect the α(0)-thalassemia mutation, which was reconfirmed using the droplet digital PCR method. Fifteen at-risk couples were identified. The timing of prenatal diagnosis was appropriate for all couples and four of the fetuses were diagnosed with Bart’s hydrops fetalis. The results were compatible with those calculated using the Hardy-Weinberg equation for a recessively inherited single gene disorder. The conclusion was that the fast-track strategy could shorten screening policy timelines, promoting early risk detection for couples and early prenatal diagnosis. The fast-track strategy might be beneficial for the prevention of hemoglobin Bart’s hydrops fetalis syndrome. 针对 α(0) 地中海贫血（东南亚和泰国类型的贫血）的早期风险检测和产前诊断，我们提出了一种快速跟踪方法：使用定量实时聚合酶链反应 (PCR)，进行直接血液 DNA 分析。血液 DNA 样本取自 1,235 对 ANC 夫妇的志愿者组。使用定量实时 PCR 对其进行评估，以检测 α(0) 地中海贫血（东南亚和泰国类型的贫血）的载体。确定危险群夫妇之后，实施羊膜穿刺术，进行进一步的产前诊断。从羊膜细胞中分离出胎儿 DNA，使用定量实时 PCR 进行特征标度，以检测 α(0) 地中海贫血突变，然后使用液滴数字 PCR 法再次确认。共确定出十五对危险群夫妇。产前诊断这一时机适用于所有夫妇，其中四例胎儿确诊为巴特水肿胎儿。我们使用 Hardy-Weinberg 方程式来计算隐性遗传单基因疾病，结果一致。结论是，此快速跟踪方法可以缩短筛选策略时间表、方便夫妇进行早期风险检测、便于早期产前诊断。此快速跟踪方法或有利于预防血红蛋白巴特水肿胎儿综合征。

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