scholarly journals Allele-specific methylated multiplex real-time quantitative PCR (ASMM RTQ-PCR), a powerful method for diagnosing loss of imprinting of the 11p15 region in Russell Silver and Beckwith Wiedemann syndromes

2011 ◽  
Vol 32 (2) ◽  
pp. 249-258 ◽  
Author(s):  
Salah Azzi ◽  
Virginie Steunou ◽  
Alexandra Rousseau ◽  
Sylvie Rossignol ◽  
Nathalie Thibaud ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Powerful Method ◽  
Real Time Quantitative Pcr ◽  
Loss Of Imprinting ◽  
Allele Specific

scholarly journals Determination of RhD Zygosity: Comparison of a Double Amplification Refractory Mutation System Approach and a Multiplex Real-Time Quantitative PCR Approach

2001 ◽  
Vol 47 (4) ◽  
pp. 667-672 ◽  
Author(s):  
Rossa W K Chiu ◽  
Michael F Murphy ◽  
Carrie Fidler ◽  
Benny C Y Zee ◽  
James S Wainscoat ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Gene Dosage ◽  
Amplification Refractory Mutation System ◽  
Pcr Assay ◽  
Real Time Quantitative Pcr ◽  
Mutation System ◽  
Allele Specific ◽  
Quantitative Pcr Assay

Abstract Background: Rh isoimmunization and hemolytic disease of the newborn still occur despite the availability of Rh immunoglobulin. For the prenatal investigation of sensitized RhD-negative pregnant women, determination of the zygosity of the RhD-positive father has important implications. The currently available molecular methods for RhD zygosity assessment, in general, are technically demanding and labor-intensive. Therefore, at present, rhesus genotype assessment is most commonly inferred from results of serological tests. The recent elucidation of the genetic structure of the prevalent RHD deletion in Caucasians, as well as the development of real-time PCR, allowed us to explore two new approaches for the molecular determination of RhD zygosity. Methods: Two methods for RhD zygosity determination were developed. The first was based on the double Amplification Refractory Mutation System (double ARMS). The second was based on multiplex real-time quantitative PCR. For the double ARMS assay, allele-specific primers were designed to directly amplify the most prevalent RHD deletion found in RhD-negative individuals in the Caucasian population. The multiplex real-time quantitative PCR assay, on the other hand, involved coamplification and quantification of RHD-specific sequences in relation to a reference gene, albumin, in a single PCR reaction. A ratio, ΔCt, based on the threshold cycle, was then determined and reflects the RHD gene dosage. Results: The allele-specific primers of the double ARMS assay reliably amplified the RHD-deleted allele and therefore accurately distinguished homozygous from heterozygous RhD-positive samples. The results were in complete concordance with serological testing. For the multiplex real-time quantitative PCR assay, the ΔCt values clearly segregated into two distinct populations according to the RHD gene dosage, with mean values of 1.70 (SD, 0.17) and 2.62 (SD, 0.29) for the homozygous and heterozygous samples, respectively (P <0.001, t-test). The results were in complete concordance with the results of serological testing as well as with the double ARMS assay. Conclusion: Double ARMS and real-time quantitative PCR are alternative robust assays for the determination of RhD zygosity.

scholarly journals Minimal Residual Disease Detection by Next-Generation Sequencing in Multiple Myeloma: A Comparison With Real-Time Quantitative PCR

2021 ◽  
Vol 10 ◽  
Author(s):  
Qiumei Yao ◽  
Yinlei Bai ◽  
Shaji Kumar ◽  
Elaine Au ◽  
Alberto Orfao ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Next Generation Sequencing ◽  
Real Time ◽  
Quantitative Pcr ◽  
Residual Disease ◽  
Next Generation ◽  
Real Time Quantitative Pcr ◽  
Allele Specific ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing

Here we compared clonotype identification by allele-specific oligonucleotide real-time quantitative-PCR (ASO RQ-PCR) and next-generation sequencing (NGS) in 80 multiple myeloma patients. ASO RQ-PCR was applicable in 49/55 (89%) and NGS in 62/78 (80%). Clonotypes identified by both methods were identical in 33/35 (94%). Sensitivity of 10−5 was confirmed in 28/29 (96%) by NGS while sensitivity of RQ-PCR was 10−5 in 7 (24%), 5 × 10−5 in 15 (52%), and 10−4 in 7 (24%). Among 14 samples quantifiable by ASO RQ-PCR, NGS yielded comparable results in 12 (86%). Applicability of NGS can be improved if immunoglobulin heavy-chain incomplete DJ primers are included.

scholarly journals Quantification and Stability of Human Adenoviruses and Polyomavirus JCPyV in Wastewater Matrices

2006 ◽  
Vol 72 (12) ◽  
pp. 7894-7896 ◽  
Author(s):  
Silvia Bofill-Mas ◽  
Nestor Albinana-Gimenez ◽  
Pilar Clemente-Casares ◽  
Ayalkibet Hundesa ◽  
Jesus Rodriguez-Manzano ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
High Stability ◽  
Wastewater Sludge ◽  
Human Polyomavirus ◽  
Sewage Effluent ◽  
Viral Contamination ◽  
Human Adenoviruses ◽  
Urban Sewage ◽  
Real Time Quantitative Pcr

ABSTRACT Human adenoviruses (HAdV) and human polyomavirus JCPyV have been previously proposed as indicators of fecal viral contamination in the environment. Different wastewater matrices have been analyzed by applying real-time quantitative PCR procedures for the presence, quantity, and stability of a wide diversity of excreted HAdV and JCPyV. High quantities of HAdV and JCPyV were detected in sewage, effluent wastewater, sludge, and biosolid samples. Both viruses showed high stability in urban sewage. These results confirm the suitability of both viruses as indicators of human fecal viral pollution.

Analysis of Caecal Microbiota in Rats Fed with Genetically Modified Rice by Real-Time Quantitative PCR

2011 ◽  
Vol 76 (1) ◽  
pp. M88-M93 ◽  
Author(s):  
Wentao Xu ◽  
Liting Li ◽  
Jiao Lu ◽  
YunBo Luo ◽  
Ying Shang ◽  
...  
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Genetically Modified ◽  
Genetically Modified Rice ◽  
Real Time Quantitative Pcr ◽  
Caecal Microbiota

scholarly journals Identification, Detection, and Enumeration of Human Bifidobacterium Species by PCR Targeting the Transaldolase Gene

2002 ◽  
Vol 68 (5) ◽  
pp. 2420-2427 ◽  
Author(s):  
Teresa Requena ◽  
Jeremy Burton ◽  
Takahiro Matsuki ◽  
Karen Munro ◽  
Mary Alice Simon ◽  
...  
Keyword(s):  
Real Time ◽  
16S Rdna ◽  
Quantitative Pcr ◽  
Gene Sequence ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Bifidobacterium Longum ◽  
Bifidobacterium Adolescentis ◽  
Fecal Samples ◽  
Real Time Quantitative Pcr ◽  
Pcr Targeting

ABSTRACT Methods that enabled the identification, detection, and enumeration of Bifidobacterium species by PCR targeting the transaldolase gene were tested. Bifidobacterial species isolated from the feces of human adults and babies were identified by PCR amplification of a 301-bp transaldolase gene sequence and comparison of the relative migrations of the DNA fragments in denaturing gradient gel electrophoresis (DGGE). Two subtypes of Bifidobacterium longum, five subtypes of Bifidobacterium adolescentis, and two subtypes of Bifidobacterium pseudocatenulatum could be differentiated using PCR-DGGE. Bifidobacterium angulatum and B. catenulatum type cultures could not be differentiated from each other. Bifidobacterial species were also detected directly in fecal samples by this combination of PCR and DGGE. The number of species detected was less than that detected by PCR using species-specific primers targeting 16S ribosomal DNA (rDNA). Real-time quantitative PCR targeting a 110-bp transaldolase gene sequence was used to enumerate bifidobacteria in fecal samples. Real-time quantitative PCR measurements of bifidobacteria in fecal samples from adults correlated well with results obtained by culture when either a 16S rDNA sequence or the transaldolase gene sequence was targeted. In the case of samples from infants, 16S rDNA-targeted PCR was superior to PCR targeting the transaldolase gene for the quantification of bifidobacterial populations.

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