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 Occurrence of Two Little Cherry Viruses in Sweet Cherry in Washington State

Plant Disease ◽  
2008 ◽  
Vol 92 (2) ◽  
pp. 234-238 ◽  
Author(s):  
N. B. Bajet ◽  
T. R. Unruh ◽  
K. L. Druffel ◽  
K. C. Eastwell
Keyword(s):  
Sweet Cherry ◽  
Prunus Avium ◽  
Washington State ◽  
Replicase Gene ◽  
Specific Primers ◽  
Reading Frame ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Affect Detection ◽  
Polymerase Chain

Little cherry disease, one of the major viral diseases of sweet cherry (Prunus avium) worldwide, is associated with either of two closteroviruses, Little cherry virus 1 (LChV-1) and Little cherry virus 2 (LChV-2). Two sets of primers corresponding to a portion of the replicase gene of LChV-1 and LChV-2 were used in one-tube reverse-transcription polymerase chain reactions to detect these viruses in total RNA extracts of field-collected sweet cherry tissues. LChV-1 and LChV-2 were detected both alone and in combination in five sweet cherry orchards in Washington State. Sequence analysis of a 240-nucleotide (nt) fragment of the replicase open reading frame (ORF)1b and a 232-nt fragment from a portion of ORF8 and the 3′ untranslated region (UTR) of LChV-1 indicated that North American (NA) isolates shared 90 to 99% nucleotide identity in both genome segments analyzed. In contrast, comparisons of NA isolates to two Eurasian isolates of LChV-1 indicated shared nucleotide identities of 79 to 82% in the replicase fragment and 89 to 90% in the ORF8/3′UTR fragment. Sequence variation in the replicase region did not affect detection of LChV-1 in 12 isolates using the replicase-specific primers reported here. This article represents the first report of LChV-1 and LChV-2 in sweet cherry in Washington.

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 ABO glycosyltransferase genotyping by polymerase chain reaction using sequence-specific primers

Blood ◽  
1996 ◽  
Vol 88 (5) ◽  
pp. 1852-1856 ◽  
Author(s):  
C Gassner ◽  
A Schmarda ◽  
W Nussbaumer ◽  
D Schonitzer
Keyword(s):  
Pcr Amplification ◽  
Abo Blood Groups ◽  
Chain Reaction ◽  
Specific Primers ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Serological Typing ◽  
Modern Molecular Biology ◽  
Specific Nucleotide Sequence ◽  
Polymerase Chain

Serological typing for the classical ABO blood groups is routinely performed using anti-A and anti-B antisera of polyclonal or monoclonal origin, which are able to distinguish four phenotypes (A, B, AB, and O). Modern molecular biology methods offer the possibility of direct ABO genotyping without the need for family investigations. Typing can be done with small amounts of DNA and without detection of blood group molecules on the surface of red blood cells. We developed a system of eight polymerase chain reactions (PCR) to detect specific nucleotide sequence differences between the ABO alleles O1, O2, A1, A2, and B. PCR amplification using sequence-specific primers and detection of amplification products by agarose gel electrophoresis is one of the fastest genotyping methods and is easy to handle. With our method we tested the A1,2BO1,2 genotypes of 300 randomly chosen persons out of a pool of platelet donors and found the results to be consistent with ABO glycosyltransferase phenotypes. We also identified a presumably new ABO allele, which may be the result of a crossing-over event between alleles O1 and A2.

scholarly journals SPATIAL-TEMPORAL DISTRIBUTION AND SEQUENCE DIVERSITY OF GROUP A HUMAN RESPIRATORY SYNCYTIAL VIRUSES IN KENYA PRECEDING THE EMERGENCE OF ON1 GENOTYPE

2021 ◽  
Author(s):  
JULIA WANGUI ◽  
David Nokes ◽  
Victor Mobegi ◽  
James Otieno ◽  
Charles Agoti ◽  
...  
Keyword(s):  
Temporal Distribution ◽  
Severe Pneumonia ◽  
Respiratory Illness ◽  
Haplotype Network ◽  
Rt Pcr ◽  
Group A ◽  
One Step ◽  
Respiratory Syncytial Viruses ◽  
Polymerase Chain ◽  
Syncytial Virus

Background: Human respiratory syncytial virus (HRSV) is a major cause of severe viral acute respiratory illness and contributes significantly to severe pneumonia cases in Africa. Little is known about its spatial-temporal distribution as defined by its genetic diversity. Methods: A retrospective study conducted utilizing archived nasopharyngeal specimens from patients attending outpatient clinics in hospitals located in five demographically and climatically distinct regions of Kenya; Coast, Western, Highlands, Eastern and Nairobi. The viral total RNA was extracted and tested using multiplex real time RT-PCR (reverse transcriptase polymerase chain reaction). A segment of the G-gene was amplified using one-step RT-PCR and sequenced by Sanger di-deoxy method. Bayesian analysis of phylogeny was utilized and subsequently median joining methods for haplotype network reconstruction. Results: Three genotypes of HRSVA were detected; GA5 (14.0%), GA2 (33.1%) and NA1 (52.9%). HRSVA prevalence varied by location from 33% to 13.2% in the Highlands and the Eastern regions respectively. The mean nucleotide diversity (Pi(π)) varied by genotype: highest of 0.018 for GA5 and lowest of 0.005 for NA1. A total of 58 haplotypes were identified (GA5 10; GA2 20; NA1 28). These haplotypes were introduced into the population locally by single haplotypes and additional subsidiary seeds amongst the GA2 and the NA1 haplotypes. Conclusions: HRSVA was found across all the regions throughout the study period and comprised three genotypes; GA5, GA2 and NA1 genotypes. The genotypes were disproportionately distributed across the regions with GA5 gradually increasing towards the Western zones and decreasing towards the Eastern zones of the country.

scholarly journals Y chromosome microdeletions in infertile male candidates for microfertilization

2008 ◽  
Vol 136 (3-4) ◽  
pp. 126-130
Author(s):  
Momcilo Ristanovic ◽  
Vera Bunjevacki ◽  
Cane Tulic ◽  
Ivana Novakovic ◽  
Tatjana Ille ◽  
...  
Keyword(s):  
Y Chromosome ◽  
Sperm Count ◽  
Gene Mutations ◽  
Control Group ◽  
Specific Primers ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Infertile Men ◽  
Y Chromosome Microdeletions ◽  
Polymerase Chain

Introduction Y chromosome microdeletions are the second most frequent genetic cause of male infertility after Klinefelter's syndrome. Objective The aim of the study was to determine the frequency of Y chromosome microdeletions in a group of infertile men with an idiophatic cause of infertility, candidates for microfertilization (Intra-cytoplasmic Sperm Injection - ICSI) in Serbia and to correlate genotype-phenotype in patients with Y chromosome microdeletions. METHOD One hundred and sixty patients with low sperm count (less than 5x106 spermatozoa/ml) were enrolled in the study. Forty patients were excluded from the study: ten because they were diagnosed with cytogenetic abnormality and thirty patients were diagnosed with other known causes of infertility. The control group consisted of 150 men who fathered at least one child in the last two years. Genomic DNA was extracted from peripheral blood samples and two multiplex polymerase chain reactions (PCR) analyses were performed using specific primers to confirm the presence or absence of Y chromosome microdeletions. Results Microdeletions were detected in 12 of 120 (10%) cases, while no deletions were detected in the control group. Of total number of 12 deletions, nine were detected in AZFc region (75%), one in AZFa (8%), and two in AZFbc (17%). Conclusion Testing for Y chromosome microdeletions should be considered as an important element in diagnosis and genetic counselling of infertile couples in Serbia. Decisions regarding the assisted reproduction should be made based on the detailed clinical, endocrinological and cytogenetic examinations, spermogram, presence or absence and type of AZF microdeletions and CFTR gene mutations. .

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 ABO glycosyltransferase genotyping by polymerase chain reaction using sequence-specific primers

Blood ◽  
1996 ◽  
Vol 88 (5) ◽  
pp. 1852-1856 ◽  
Author(s):  
C Gassner ◽  
A Schmarda ◽  
W Nussbaumer ◽  
D Schonitzer
Keyword(s):  
Pcr Amplification ◽  
Abo Blood Groups ◽  
Chain Reaction ◽  
Specific Primers ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Serological Typing ◽  
Modern Molecular Biology ◽  
Specific Nucleotide Sequence ◽  
Polymerase Chain

Abstract Serological typing for the classical ABO blood groups is routinely performed using anti-A and anti-B antisera of polyclonal or monoclonal origin, which are able to distinguish four phenotypes (A, B, AB, and O). Modern molecular biology methods offer the possibility of direct ABO genotyping without the need for family investigations. Typing can be done with small amounts of DNA and without detection of blood group molecules on the surface of red blood cells. We developed a system of eight polymerase chain reactions (PCR) to detect specific nucleotide sequence differences between the ABO alleles O1, O2, A1, A2, and B. PCR amplification using sequence-specific primers and detection of amplification products by agarose gel electrophoresis is one of the fastest genotyping methods and is easy to handle. With our method we tested the A1,2BO1,2 genotypes of 300 randomly chosen persons out of a pool of platelet donors and found the results to be consistent with ABO glycosyltransferase phenotypes. We also identified a presumably new ABO allele, which may be the result of a crossing-over event between alleles O1 and A2.

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