scholarly journals New methods as alternative or corrective measures for the pitfalls and artifacts of reverse transcription and polymerase chain reactions (RT-PCR) in cloning chimeric or antisense-accompanied RNA

RNA Biology ◽  
2013 ◽  
Vol 10 (6) ◽  
pp. 957-967 ◽  
Author(s):  
Chengfu Yuan ◽  
Yongming Liu ◽  
Min Yang ◽  
D. Joshua Liao
Keyword(s):  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
New Methods ◽  
Corrective Measures ◽  
Polymerase Chain

scholarly journals Characteristic Differences in Reverse Transcription-Polymerase Chain Reaction Products of Ovine, Bovine, and Human Respiratory Syncytial Viruses

1993 ◽  
Vol 5 (3) ◽  
pp. 322-328 ◽  
Author(s):  
Richard D. Oberst ◽  
Michael P. Hays ◽  
Jim F. Evermann ◽  
Clayton L. Kelling
Keyword(s):  
Reverse Transcription ◽  
Reaction Products ◽  
Rt Pcr ◽  
Specific Primers ◽  
F Protein ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Respiratory Syncytial Viruses ◽  
Polymerase Chain ◽  
Syncytial Virus

In reverse transcription-polymerase chain reactions (RT-PCR) and DNA hybridizations using primers and an oligonucleotide probe to the fusion (F) protein mRNA of bovine respiratory syncytial virus (BRSV), all the BRSV isolates and a goat isolate could be distinguished from prototype isolates of human respiratory syncytial viruses (HRSV) and ovine (sheep and bighorn sheep) respiratory syncytial viruses (RSV). However, RT-PCR amplifications with primers to sequences of the HRSV F protein mRNA resulted in amplified products of ≍ 243 bp if mRNA templates of subgroup A HRSV strains were present and slightly larger amplified products with subgroup B HRSV strains. No amplified products were observed in HRSV-primed RT-PCR with BRSV or goat or ovine RSV mRNA templates. Although the ovine RSV isolates were antigenically cross-reactive with the goat RSV, HRSV and BRSV isolates, they were not amplified with either HRSV- or BRSV-specific primers in RT-PCR. These results confirm previous immunological comparisons suggesting that some ovine RSV isolates should be considered as distinct respiratory syncytial viruses.

scholarly journals SARS-CoV-2 show no infectivity at later stages in a prolonged COVID-19 patient despite positivity in RNA testing

2021 ◽  
Author(s):  
Xiu-Feng Wan ◽  
Cynthia Y Tang ◽  
Detlef Ritter ◽  
Yang Wang ◽  
Tao Li ◽  
...  
Keyword(s):  
Large Scale ◽  
Disease Onset ◽  
Nasopharyngeal Swab ◽  
Rt Pcr ◽  
Virus Growth ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Asymptomatic Patients ◽  
Hospitalization Period ◽  
Polymerase Chain

Inpatient COVID-19 cases present enormous costs to patients and health systems. Many hospitalized patients may still test COVID-19 positive, even after resolution of symptoms. Thus, a pressing concern for clinicians is the safety of discharging these asymptomatic patients if they have any remaining infectivity. This case report explores the viral viability in a patient with persistent COVID-19 over the course of a two-month hospitalization. Positive nasopharyngeal swab samples, analyzed by quantitative reverse transcription polymerase chain reactions (qRT-PCR), were collected and isolated in the laboratory, and infectious doses were analyzed throughout the hospitalization period. The patient experienced waning symptoms by hospital day 40 and had no viable virus growth in the laboratory by hospital day 41, suggesting no risk of infectivity, despite positive RT-PCR results, which prolonged his hospital stay. Notably, this case showed infectivity for at least 24 days from disease onset, which is longer than the discontinuation of transmission-based precautions recommendation by CDC. Thus, our findings suggest that the timeline for discontinuing transmission-based precautions may need to be extended for patients with prolonged illness. Additional large-scale studies are needed to draw definitive conclusions on the appropriate clinical management for these patients.

scholarly journals Prognostic value of Serum miR-217 Level in Osteosarcoma Patients

2020 ◽  
Author(s):  
Jia Ye ◽  
Zhi-Hui Jin ◽  
Ren Chen ◽  
Sen Chen ◽  
Yi-Jun Ren ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Poor Prognosis ◽  
Prognostic Significance ◽  
Serum Levels ◽  
High Serum ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Low Serum

Abstract Background MicroRNA (miR)-217 is a tumor suppressor significantly associated with osteosarcoma. We try to evaluate serum levels miR-217 in osteosarcoma patients and evaluate its prognostic significance. Methods A total of 163 consecutive osteosarcoma patients and 96 healthy participates were enrolled. Serum miR-217 levels were evaluated by using real-time quantitative reverse transcription polymerase chain reactions(RT-PCR). The association between serum miR-217 level and survival outcomes was evaluated by univariate and multivariate analysis. Results Serum miR-217 levels in osteosarcoma patients was significantly lower than healthy volunteers (P<0.05). Low serum miR-217 was significantly related to advanced cancer and metastasis (both P<0.05). Moreover, patients with a low serum miR-217 had a poorer overall survival than those with a high serum miR-217 levels (P<0.05). Serum miR-217 level also been showed as independent risk factor for osteosarcoma in multivariate analysis (HR, 0.42; 95%CI: 0.12–0.98; p<0.01). Conclusions Serum miR-217 levels was significantly downregulated in osteosarcoma patients and remarkably associated with poor prognosis, indicating that serum miR-217 might serve as a useful diagnostic and prognostic indictor for osteosarcoma.

scholarly journals Prognostic value of Serum miR-217 Level in Osteosarcoma Patients

2021 ◽  
Author(s):  
Jia Ye ◽  
Zhi-Hui Jin ◽  
Ren Chen ◽  
Sen Chen ◽  
Yi-Jun Ren ◽  
...  
Keyword(s):  
Multivariate Analysis ◽  
Prognostic Significance ◽  
Serum Levels ◽  
Prognostic Indicator ◽  
High Serum ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain ◽  
Low Serum

Abstract Background: MicroRNA (miR)-217 is a tumor suppressor significantly associated with osteosarcoma. We try to evaluate serum levels miR-217 in osteosarcoma patients and evaluate its prognostic significance.Methods: A total of 163 consecutive osteosarcoma patients and 96 healthy participates were enrolled. Serum miR-217 levels were evaluated by using real-time quantitative reverse transcription polymerase chain reactions(RT-PCR). The association between serum miR-217 level and survival outcomes was evaluated by univariate and multivariate analysis. Results: Serum miR-217 levels in osteosarcoma patients was significantly lower than healthy volunteers (P<0.05). Low serum miR-217 was significantly related to advanced cancer and metastasis (both P<0.05). Moreover, patients with a low serum miR-217 had a poorer overall survival than those with a high serum miR-217 levels (P<0.05). Serum miR-217 level also been showed as independent risk factor for osteosarcoma in multivariate analysis (HR, 0.42; 95%CI: 0.12–0.98; p<0.01). Conclusions: Serum miR-217 levels was significantly downregulated in osteosarcoma patients and remarkably associated with poor prognosis, indicating that serum miR-217 might serve as a useful diagnostic and prognostic indicator for osteosarcoma.

Identification of the NO Synthase isoforms Expressed in Human Neutrophil Granulocytes, Megakaryocytes and Platelets

1997 ◽  
Vol 77 (01) ◽  
pp. 163-167 ◽  
Author(s):  
Thomas Wallerath ◽  
Ingolf Gath ◽  
Walter E Aulitzky ◽  
Jennifer S Pollock ◽  
Hartmut Kleinert ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Human Platelets ◽  
No Synthase ◽  
Immunofluorescent Staining ◽  
Human Neutrophils ◽  
Neutrophil Granulocytes ◽  
Rt Pcr ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Polymerase Chain

SummaryUsing Western blot and fluorescent immunocytochemistry, NOS III (or ecNOS) and NOS II (or iNOS), but no NOS I (or ncNOS), were identified in preparations of human platelets. Reverse-transcription polymerase chain reactions (RT-PCR) demonstrated NOS III mRNA, but no NOS II mRNA (which is short-lived) and no NOS I mRNA in platelets. Immunofluorescent staining of human bone marrow smears showed the presence of NOS III, but not NOS I in megakaryocytes. A subpopulation of megakaryocytes also expressed NOS II. In preparations of human neutrophils, immunocytochemistry demonstrated NOS I in all cells, whereas no NOS III was detected. The few NOS II positive cells were characterized as contaminating eosinophils. Similarly, in RT-PCR, transcripts for NOS I and NOS II, but not for NOS III, were identified. Thus, the constitutive NOS isoform in megakaryocytes and platelets is NOS III, whereas neutrophils express NOS I. Some megakaryocytes and eosinophils also express NOS II.

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