Establishment of a cell‐based quantitative reverse transcription‐polymerase chain reaction (RT‐qPCR) assay for detection of multivalent rotavirus vaccine

2020 ◽  
Vol 92 (12) ◽  
pp. 3157-3164
Author(s):  
Yunjin Wang ◽  
Yueyue Liu ◽  
Mingqiang Wang ◽  
Li Yu ◽  
Chao Ma ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Qpcr Assay ◽  
Rotavirus Vaccine ◽  
Chain Reaction ◽  
A Cell ◽  
Polymerase Chain

Junctions of the AML1/MTG8(ETO) fusion are constant in t(8;21) acute myeloid leukemia detected by reverse transcription polymerase chain reaction

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1270-1276 ◽  
Author(s):  
T Kozu ◽  
H Miyoshi ◽  
K Shimizu ◽  
N Maseki ◽  
Y Kaneko ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Myeloid Leukemia ◽  
Cell Line ◽  
Reverse Transcription ◽  
Myeloid Leukemia ◽  
Chain Reaction ◽  
Translocation Breakpoints ◽  
A Cell ◽  
Polymerase Chain ◽  
Acute Myeloid

Abstract The chromosomal translocation, t(8;21), is found frequently in acute myeloid leukemia (AML) with maturation (FAB-M2). We have previously mapped the translocation breakpoints of t(8;21) in a specific intron of the AML1 gene on chromosome 21. In this study, we cloned cDNAs synthesized from a cell line carrying t(8;21) by reverse transcription polymerase chain reaction (RT-PCR) using an AML1-specific primer. The analysis of the cDNAs structure has led to the identification of the fusion of AML1 with a gene named MTG8 on chromosome 8, which seems to be identical to ETO. Northern analysis using MTG8 (ETO) probes detected 7.8-kb and 6.2-kb RNAs and several minor RNAs in the cell line with t(8;21), but failed to detect any transcripts in a cell line without t(8;21). A set of primers were designed to detect the AML1/MTG8(ETO) fusion by PCR. The PCR amplified identical products in all 6 patients and one cell line with t(8;21), suggesting that the AML1/MTG8(ETO) fusion is a constant feature associated with t(8;21) and the junctions of the AML1/MTG8(ETO) fusion are restricted in a unique site. Because the PCR detection of the AML1/MTG8(ETO) fusion at the RNA level is highly sensitive, it can be used as a sensitive method for diagnosis and detection of minimal residual disease in t(8;21) leukemia.

scholarly journals Clinical Validation of a Novel Commercial Reverse Transcription–Quantitative Polymerase Chain Reaction Screening Assay for Detection of ALK Translocations and Amplifications in Non–Small Cell Lung Carcinomas

2015 ◽  
Vol 140 (7) ◽  
pp. 690-693 ◽  
Author(s):  
Chunyan Liu ◽  
Kristi Pepper ◽  
Heather Hendrickson ◽  
Philip T. Cagle ◽  
Bryce P. Portier
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Quantitative Polymerase Chain Reaction ◽  
Genetic Alterations ◽  
Qpcr Assay ◽  
Egfr Mutations ◽  
Chain Reaction ◽  
Lung Cancers ◽  
Anaplastic Lymphoma ◽  
Polymerase Chain

Context.— EGFR mutations and anaplastic lymphoma kinase (ALK) translocations have significant biologic and therapeutic implications in lung cancers, particularly lung adenocarcinomas. ALK translocations are less frequent compared with EGFR mutations; interestingly, these two abnormalities are most commonly mutually exclusive. The 2013 College of American Pathologists/Association for Molecular Pathology/International Association for the Study of Lung Cancer molecular testing guideline for lung cancers recommend a testing algorithm in which detection of ALK translocations using fluorescence in situ hybridization (FISH) is to be performed following testing for EGFR mutations. Such an algorithm is cost-effective but potentially slows down turnaround time; and as a secondary test, ALK FISH assay may not be completed because it requires the use of additional tissue, and the small biopsies or cytology specimens may have been exhausted in the extraction of nucleic acid for EGFR mutation screening.Objective.—To provide efficient testing of both EGFR and ALK genetic alterations in small biopsies and cytology specimens.Design.—We validated a highly sensitive ALK reverse transcription–quantitative polymerase chain reaction (RT-qPCR) assay as a screening tool for ALK translocations and amplifications.Results.—We performed a retrospective review of cases previously tested by FISH and found that all FISH ALK translocation–positive specimens were RT-qPCR positive, and all FISH ALK translocation–negative cases were RT-qPCR negative (the sensitivity and specificity of the ALK RT-qPCR assay were 100%).Conclusion.—This assay allows rapid identification of ALK alterations, can be performed in conjunction with EGFR testing, and does not require use of valuable additional tumor tissue.

scholarly journals An Ultrafast One-Step Quantitative Reverse Transcription–Polymerase Chain Reaction Assay for Detection of SARS-CoV-2

2021 ◽  
Vol 12 ◽  
Author(s):  
Jadranka Milosevic ◽  
Mengrou Lu ◽  
Wallace Greene ◽  
Hong-Zhang He ◽  
Si-Yang Zheng
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Polymerase Chain Reaction Assay ◽  
Qpcr Assay ◽  
Nasopharyngeal Swab ◽  
Chain Reaction ◽  
Running Time ◽  
Clinical Sensitivity ◽  
One Step ◽  
Polymerase Chain

We developed an ultrafast one-step RT-qPCR assay for SARS-CoV-2 detection, which can be completed in only 30 min on benchtop Bio-Rad CFX96. The assay significantly reduces the running time of conventional RT-qPCR: reduced RT step from 10 to 1 min, and reduced the PCR cycle of denaturation from 10 to 1 s and extension from 30 to 1 s. A cohort of 60 nasopharyngeal swab samples testing showed that the assay had a clinical sensitivity of 100% and a clinical specificity of 100%.

scholarly journals Junctions of the AML1/MTG8(ETO) fusion are constant in t(8;21) acute myeloid leukemia detected by reverse transcription polymerase chain reaction

Blood ◽  
1993 ◽  
Vol 82 (4) ◽  
pp. 1270-1276 ◽  
Author(s):  
T Kozu ◽  
H Miyoshi ◽  
K Shimizu ◽  
N Maseki ◽  
Y Kaneko ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Myeloid Leukemia ◽  
Cell Line ◽  
Reverse Transcription ◽  
Myeloid Leukemia ◽  
Chain Reaction ◽  
Translocation Breakpoints ◽  
A Cell ◽  
Polymerase Chain ◽  
Acute Myeloid

The chromosomal translocation, t(8;21), is found frequently in acute myeloid leukemia (AML) with maturation (FAB-M2). We have previously mapped the translocation breakpoints of t(8;21) in a specific intron of the AML1 gene on chromosome 21. In this study, we cloned cDNAs synthesized from a cell line carrying t(8;21) by reverse transcription polymerase chain reaction (RT-PCR) using an AML1-specific primer. The analysis of the cDNAs structure has led to the identification of the fusion of AML1 with a gene named MTG8 on chromosome 8, which seems to be identical to ETO. Northern analysis using MTG8 (ETO) probes detected 7.8-kb and 6.2-kb RNAs and several minor RNAs in the cell line with t(8;21), but failed to detect any transcripts in a cell line without t(8;21). A set of primers were designed to detect the AML1/MTG8(ETO) fusion by PCR. The PCR amplified identical products in all 6 patients and one cell line with t(8;21), suggesting that the AML1/MTG8(ETO) fusion is a constant feature associated with t(8;21) and the junctions of the AML1/MTG8(ETO) fusion are restricted in a unique site. Because the PCR detection of the AML1/MTG8(ETO) fusion at the RNA level is highly sensitive, it can be used as a sensitive method for diagnosis and detection of minimal residual disease in t(8;21) leukemia.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain
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