scholarly journals The occurrence of the Tulip breaking virus in tulips in the northern part of Turkey

2019 ◽  
Vol 31 (2) ◽  
pp. 263-268
Author(s):  
Ilyas Deligoz ◽  
Mehmet Ali Sevik
Keyword(s):  
Viral Disease ◽  
Cut Flower ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Black Sea Region ◽  
Tulip Breaking Virus ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Garden Plant ◽  
Das Elisa

AbstractThe tulip (Tulipa sp.) is one of the most important ornamental bulbous plants, which has been cultivated as a cut-flower, potted, and garden plant, and used for landscaping in Turkey. This study investigated the occurrence of a viral disease in the tulip cultivars Strong Gold, Pretty Woman and Purple Prince that causes striping of the leaves, flames of different colours on the petals and mosaic patterns on the leaves, in Samsun province of Turkey. Surveys of virus-infected tulip plants were carried out in the Middle Black Sea Region of Turkey in 2015-2016. A total of 212 samples were collected from four locations and checked by biological, serological and molecular methods for the presence of the Tulip breaking virus (TBV). TBV was detected in the leaves and flowers by double-antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) in the tulip cultivars (15.5%) tested from Samsun province. TBV infection was found at the highest rate in the cultivar Strong Gold (19.7%), followed by Pretty Woman (14.1%) and Purple Prince (12.8%). The presence of TBV in samples was further confirmed by reverse transcription polymerase chain reaction (RT-PCR) assays. This is the first report on TBV naturally infecting tulips in Samsun province, Turkey.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

scholarly journals The Value of Pre-operative Testing for COVID-19 in Urological Patients

2020 ◽  
Vol 3 (3) ◽  
pp. e29-e34
Author(s):  
Vasileios Bonatsos ◽  
Asif Raza
Keyword(s):  
Gold Standard ◽  
Diagnostic Tests ◽  
Acute Infection ◽  
World Health Organisation ◽  
World Health ◽  
Rt Pcr ◽  
Chain Reaction ◽  
The World ◽  
Polymerase Chain ◽  
Pcr Assays

According to the World Health Organisation there have been 30,055,710 confirmed COVID-19 cases and 933,433 confirmed deaths across 216 countries globally. The availability of the complete SARS-CoV-2 genome relatively early in the epidemic has enabled the development of tests for the diagnosis of COVID-19. There are two broad categories of SARS-CoV-2 diagnostic tests currently in use or development: (1) Real-time reverse transcriptase polymerase chain reaction (RT-PCR) tests and (2) serology tests. RT-PCR is considered the gold standard and preferred method of diagnosis of acute infection. There is, however, a plethora of laboratory-developed and commercial RT-PCR assays with different gene targets. We discuss the value of pre-operative testing for COVID-19 before urological surgery.

Elucidating the Pivotal Role of Immune Players in the Management of COVID-19: Focus on Mesenchymal Stem Cells and Inflammation

2020 ◽  
Vol 15 ◽  
Author(s):  
Seidu A. Richard ◽  
Sylvanus Kampo ◽  
Marian Sackey ◽  
Maite Esquijarosa Hechavarria ◽  
Alexis D. B. Buunaaim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Disease Burden ◽  
Viral Disease ◽  
Inflammatory Factors ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Paracrine Secretion ◽  
Polymerase Chain

: The world is currently engulfed with a viral disease with no cure. So, far, millions of people are infected with the virus across the length and breadth of world with thousand losing their lives each passing day. The WHO is February 2020 classified the virus as a coronavirus and the name Coronavirus-19 (CoV-19) was offered to the virus. The disease caused by the virus was termed coronavirus disease-19 (COVID-19). The pathogenesis of COVID-19 is associated with elevation of several immune plays as well as inflammatory factors which contributes to cytokine storms. Currently, the detection of CoV-19 RNA is through reverse transcriptase polymerase chain reaction (RT-PCR). Mesenchymal stem cells (MSCs) are capable of suppressing several kinds of cytokines via the paracrine secretion system. Therefore, MSCs therapy could be game charges in the treatment of the current COVID-19 pandemic. Also, intravenous IG may be capable of suppressing the high expression of IL-6 by the CoV-19 resulting in lessen disease burden. Anti-inflammatory medications like, corticosteroids, tocilizumab, glycyrrhetinic acid, as well as etoposide may be very advantageous in decreasing the COVID-19 burden because, their mode of action targets the cytokine storms initiated by the CoV-19. It is important to indicate that, these medication does not target the virus itself. Therefore, potent CoV-19 anti-viral medications are needed to completely cure patients with COVID-19. Also, a vaccine is urgently needed to stop the spread of the virus. This review therefore elucidates the immune players in the management of COVID-19; focusing principally on MSCs and inflammatory mediators.

scholarly journals Molecular analysis of t(11;19) breakpoints in childhood acute leukemias

Blood ◽  
1996 ◽  
Vol 87 (11) ◽  
pp. 4804-4808 ◽  
Author(s):  
JE Rubnitz ◽  
FG Behm ◽  
AM Curcio-Brint ◽  
RP Pinheiro ◽  
AJ Carroll ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Myeloid Leukemia ◽  
Lymphoblastic Leukemia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Acute Leukemias ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Childhood Acute Leukemia ◽  
Acute Myeloid

MLL is fused to ENL or ELL in acute leukemias that contain t(ll;19)(q23;p13). Although ENL and ELL localize to chromosome 19, bands p13.3 and p13.1, respectively, these breakpoints are not always readily distinguished by standard cytogenetics. We therefore used reverse transcriptase-polymerase chain reaction (RT-PCR) assays to analyze 26 cases of childhood acute leukemia containing t(11;19) to determine the frequencies of ENL and ELL involvement. All 17 cases of acute lymphoblastic leukemia (ALL) had MLL/ENL fusion transcripts. By contrast, of the 9 cases of acute myeloid leukemia (AML) analyzed, 6 had MLL/ENL fusions, 2 had MLL/ELL fusions, and 1 case had no RT-PCR- detectable MLL fusion mRNA. These data suggest that the majority of 11;19 translocations involve ENL, whereas involvement of ELL is relatively uncommon in childhood acute leukemia and may be restricted to AML.

scholarly journals Molecular (real-time reverse transcription polymerase chain reaction) diagnosis of SARS-CoV-2 infections: complexity and challenges

2021 ◽  
Vol 45 (3) ◽  
pp. 135-142
Author(s):  
Shneh Sethi ◽  
Trinad Chakraborty
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Reverse Transcription ◽  
Fatal Outcome ◽  
Respiratory Illness ◽  
World Health ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Pcr Assays

Abstract The outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first recorded in Wuhan, China. The World Health Organization initially classified COVID-19 as a public health emergency and subsequently declared the disease a global pandemic. COVID-19 can take at least three distinct forms: severe acute distress syndrome with a potentially fatal outcome, mild respiratory illness (pneumonia with eventual recovery) and asymptomatic infection. All three disease forms have the potential to transmit the infection to healthy contacts. At present, real-time reverse transcription polymerase chain reaction (RT-PCR) is the only available laboratory tool to confirm the presence of viral RNA in patient specimens. These assays are designed to detect one or more (at least 2) SARS-CoV-2 RNA gene targets allowing the detection of the virus. Commercially available RT-PCR assays employ various gene targets of the viral genome in their assay systems. Additionally, there are differences in primer selection for the same gene region of SARS-CoV-2. At present, it is unclear whether the results from different RT-PCR assays are comparable in detecting the spectrum of COVID-19 manifestations. The purpose of the present article is twofold: first, to briefly focus on the findings of these reports; and second, to emphasize the various challenges and flaws that can potentially impact the diagnostic accuracy of RT-PCR testing for SARS-CoV-2.

scholarly journals Estimating false-negative detection rate of SARS-CoV-2 by RT-PCR

2020 ◽  
Author(s):  
Paul Wikramaratna ◽  
Robert S Paton ◽  
Mahan Ghafari ◽  
José Lourenço
Keyword(s):  
Symptom Onset ◽  
Detection Rate ◽  
False Negative ◽  
Rt Pcr ◽  
Chain Reaction ◽  
False Negatives ◽  
Clinical Stages ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Negative Detection

AbstractReverse transcription-polymerase chain reaction (RT-PCR) assays are used to test patients and key workers for infection with the causative SARS-CoV-2 virus. RT-PCR tests are highly specific and the probability of false positives is low, but false negatives can occur if the sample contains insufficient quantities of the virus to be successfully amplified and detected. The amount of virus in a swab is likely to vary between patients, sample location (nasal, throat or sputum) and through time as infection progresses. Here, we analyse publicly available data from patients who received multiple RT-PCR tests and were identified as SARS-CoV-2 positive at least once. We identify that the probability of a positive test decreases with time after symptom onset, with throat samples less likely to yield a positive result relative to nasal samples. Empirically derived distributions of the time between symptom onset and hospitalisation allowed us to comment on the likely false negative rates in cohorts of patients who present for testing at different clinical stages. We further estimate the expected numbers of false negative tests in a group of tested individuals and show how this is affected by the timing of the tests. Finally, we assessed the robustness of these estimates of false negative rates to the probability of false positive tests. This work has implications both for the identification of infected patients and for the discharge of convalescing patients who are potentially still infectious.

scholarly journals Inter-laboratory quality control for hormone-dependent gene expression in human breast tumors using real-time reverse transcription-polymerase chain reaction

2004 ◽  
Vol 11 (3) ◽  
pp. 489-495 ◽  
Author(s):  
P de Cremoux ◽  
I Bieche ◽  
C Tran-Perennou ◽  
S Vignaud ◽  
E Boudou ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Quality Control ◽  
Real Time ◽  
Target Genes ◽  
Correlation Coefficients ◽  
Breast Tumors ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Pcr Assays

Quantitative reverse transcription-polymerase chain reaction (RT-PCR) used to detect minor changes in specific mRNA concentrations may be associated with poor reproducibility. Stringent quality control is therefore essential at each step of the protocol, including the PCR procedure. We performed inter-laboratory quality control of quantitative PCR between two independent laboratories, using in-house RT-PCR assays on a series of hormone-related target genes in a retrospective consecutive series of 79 breast tumors. Total RNA was reverse transcribed in a single center. Calibration curves were performed for five target genes (estrogen receptor (ER)α, ERβ, progesterone receptor (PR), CYP19 (aromatase) and Ki 67) and for two reference genes (human acidic ribosomal phosphoprotein PO (RPLPO) and TATA box-binding protein (TBP)). Amplification efficiencies of the calibrator were determined for each run and used to calculate mRNA expression. Correlation coefficients were evaluated for each target and each reference gene. A good correlation was observed for all target and reference genes in both centers using their own protocols and kits (P < 0.0001). The correlation coefficients ranged from 0.90 to 0.98 for the various target genes in the two centers. A good correlation was observed between the level of expression of the ERα and the PR transcripts (P < 0.001). A weak inverse correlation was observed in both centers between ERα and ERβ levels, but only when TBP was the reference gene. No other correlation was observed with other parameters. Real-time PCR assays allow convenient quantification of target mRNA transcripts and quantification of target-derived nucleic acids in clinical specimens. This study addresses the importance of inter-laboratory quality controls for the use of a panel of real-time PCR assays devoted to clinical samples and protocols and to ensure their appropriate accuracy. This can also facilitate exchanges and multicenter comparison of data.

scholarly journals Role of HRCT Scan of Chest in the Evaluation of COVID-19 Pneumonia

2021 ◽  
Vol 34 (1) ◽  
pp. 109-114
Author(s):  
Md Hafizur Rahman ◽  
Nashid Amir ◽  
Md Anisur Rahman ◽  
AHM Tohurul Islam ◽  
Md Saiful Islam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
High Specificity ◽  
Viral Disease ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Severity Scoring ◽  
Proper Diagnosis ◽  
Disease Spreading ◽  
Polymerase Chain

Coronavirus disease (COVID-19) pneumonia emerged in Wuhan, China, in December 2019. It is a highly contagious viral disease spreading worldwide, with a rapid increase in the number of cases & deaths. COVID-19 pneumonia is characterized by fever, fatigue, dry cough, and dyspnea with other systemic features such as diarrhea, altered sensorium, stroke & multi-organ failure. HRCT chest is one of the most sensitive modalities for early detection of COVID-19 pneumonia & monitor the outcome of these patients. It is an important complement to the reverse transcriptase polymerase chain reaction (RT-PCR) tests. HRCT shows high specificity & sensitivity in detection of COVID-19 pneumonia being 90.7% & 70.8% respectively. In this pandemic situation, proper diagnosis & management of COVID-19 positive cases largely depends on HRCT findings & severity scoring. TAJ 2021; 34: No-1: 109-114

scholarly journals Occurrence of Maize rayado fino virus in Maize in Argentina

Plant Disease ◽  
2000 ◽  
Vol 84 (9) ◽  
pp. 1046-1046 ◽  
Author(s):  
M. P. Giménez-Pecci ◽  
E. Oliveira ◽  
R. Resende ◽  
C. Borgogno ◽  
C. F. Nome ◽  
...  
Keyword(s):  
Enzyme Linked Immunosorbent Assay ◽  
Plant Tissues ◽  
Rt Pcr ◽  
Xylem Parenchyma ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Maize Rayado Fino Virus ◽  
Size Relation ◽  
Pcr Conditions ◽  
Das Elisa

Symptoms of fine chlorotic stipple-striping of the veins, chlorosis, numerous dots and stripes, formation of holes in the leaf blade, and ears reduced in size, bearing few grains, were observed in maize crops in Tafí del Valle (Tucumán Province), Orán, El Galpón (Salta Province), Tilcara and Yaví (Jujuy Province), the subtropical area of northwest Argentina where the leafhopper vector Dalbulus maidis (DeLong & Wolcott) is present. Maize rayado fino virus (MRFV) was detected in these samples by a positive reaction in double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) using an AGDIA kit. Electron microscopy revealed abundant isometric particles about 30 nm in diameter in the cytoplasm and vacuoles of phloem cells and xylem parenchyma cells. The virus was also detected by reverse transcription polymerase chain reaction (RT-PCR) using a primer pair MRFV-09/MRFV-10. Primers and PCR conditions were as previously described (1). Virus amplification was observed only in samples from symptomatic plants. In 1981 (2), the presence of MRFV in Argentina was revealed by serological assay in plants from temperate central areas. No further reports were released since then. This is the first evidence of MRFV in subtropical areas of Argentina and identification of the virus by combining DAS-ELISA, particle size, relation with plant tissues, and RTPCR. References: (1) R. W. Hammond et al. J. Gen. Virol. 78:3153, 1997. (2) S. F. Nome et al. RIA XIX:257, 1984.

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