Abstract 43: Real Time Polymerase Chain Reaction Assays For Detection Of Respiratory Viruses In Kawasaki Disease Patients

Circulation ◽  
2015 ◽  
Vol 131 (suppl_2) ◽  
Author(s):  
Young Mi Hong ◽  
Seul Bee Lee ◽  
Han Seul Choi ◽  
Jung Hyun Kwon ◽  
Hae Soon Kim ◽  
...  

Purpose: Respiratory symptoms are frequently observed in children with Kawasaki disease (KD) during the acute phase. The association rate of KD with antecedent respiratory illness has been reported to range from 56 to 83%. Clinical and epidemiologic features of KD support an infectious cause, but the etiology remains unknown. We investigated the association of respiratory viruses in children with KD using multiplex reverse transcriptase-polymerase chain reaction (RT-PCR). Methods: 138 KD patients were enrolled from January 2010 to June 2013. Two study groups (Group 1; n=94, KD without respiratory symptoms, Group 2; n=44, KD with respiratory symptoms) were compared with a control group (Group 3; n=5, febrile patients with respiratory symptoms). Laboratory data were obtained from each patient including complete blood count (CBC), erythrocyte sedimentation rate (ESR), platelet count, alanine aminotransferase (ALT), aspartate aminotransferase (AST), serum total protein, albumin, C-reactive protein (CRP), NT-pro brain natriuretic peptide (BNP). Echocardiographic measurements were compared between the three groups. RT-PCR was performed using nasopharyngeal secretion to screen for the presence of 14 viruses (corona virus, parainfluenza virus 1, 2 and 3, influenza A and B, respiratory syncytial virus A and B, rhino virus A, B and C, metapneumo virus, adenovirus, and bocavirus) in groups 2 and 3. Results: The rate of KD with respiratory symptoms was 17.5%. The duration of fever was significantly longer and coronary artery diameter was significantly larger in group 2 than in group 1. Coronary artery diameter, CRP, platelet count, ALT, and NT-pro BNP were significantly higher and albumin lower in group 2 compared with group 3. Detection rate of adenovirus was 55.0% in group 2 and 28.6% in group 3. Conclusion: A positive RT-PCR for respiratory viruses may be helpful to elucidate the specific virus in KD patients with respiratory symptoms. NT-proBNP is a very important diagnostic tool in differentiating KD from other febrile viral respiratory infaction.

2020 ◽  
Vol 65 (12) ◽  
pp. 767-770
Author(s):  
Olga Gennadjevna Kimirilova ◽  
G. A. Kharchenko

The urgency of the problem of coxyellosis in children is determined by the endemic nature of this pathology for a number of regions of Russia. The purpose of the study: to evaluate the results of diagnosis of coxyellosis in children using the methods of complement binding reaction (RSC), enzyme immunoassay (ELISA), and polymerase chain reaction (PCR). Retrospective analysis of the survey on Coxiella in 3 groups of children aged 7 to 17 years: group 1 (n=30) method RSK; group 2 (n=34) - by ELISA; group 3 (n=35) - PCR, were hospitalized in GBUZ «Regional clinical infectious hospital named. A. M. Nicholi» Astrakhan in the period from January 2010 to January 2020. The most informative methods of diagnosis of coxyellosis in children during the first 7 days from the onset of the disease is the PCR reaction (specificity-94%, sensitivity-91%), after the 7th day of the disease ELISA (specificity -91%, sensitivity - 94%).The sensitivity of the RSC method is 70%, the specificity is 87%.


2018 ◽  
Vol 68 (3) ◽  
pp. 299
Author(s):  
Ο. ASLAN ◽  
Α. GENCAY GOKSU ◽  
Ν. APAYDIN

This study was carried out to measure the plasma malondialdehyde (MDA) level, and erythrocyte glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) activities in lambs with Pestivirus. Sixty lambs aged between 2.5-3 months old were included in the study. Blood and faeces samples were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) test, using pan-Pestivirus primers. Clinical and virological examinations revealed that the numbers of non-infected and Pestivirus -infected animals were 20 (Group 1) and 40 (Group 2), respectively. Plasma MDA levels were found significantly higher in lambs with Pestivirus than that of controls (p<0.05) whilst the erythrocyte GSH-Px, CAT and SOD activities were significantly lower in lambs with Pestivirus (p<0.05) than in the controls. The determination of increased plasma MDA levels and decreased erythrocyte GSH-Px, CAT and SOD activities in the infected lambs confirms that oxidative stress is observed in Pestivirus. The presence of oxidative stress in Pestivirus indicates that the equilibrium between oxidants and antioxidants is shifted towards oxidants in lambs with Pestivirus.


2003 ◽  
Vol 112 (3) ◽  
pp. 252-257 ◽  
Author(s):  
Toshio Ishibashi ◽  
Hiroko Monobe ◽  
Masanobu Shinogami ◽  
Yuka Nomura ◽  
Jun Yano

Because respiratory viruses play an important role in the causation and pathogenesis of acute otitis media (AOM), determining which virus has infected a child is important with respect to vaccines and antiviral drugs. In some instances, this information might be used to prevent the occurrence of AOM. We used a rapid, economical, and sensitive diagnostic system involving a multiplex nested reverse transcription–polymerase chain reaction (RT-PCR) assay to detect various respiratory viruses in clinical specimens of middle ear fluid (MEF) from children with AOM in our hospital. Multiplex RT-PCR was completed on 40 MEF samples from 28 infants and children less than 6 years old with AOM. Viral RNA was detected in 17 MEF samples (43%). Respiratory syncytial virus type A was present in 12 samples, adenovirus in 3, rhinovirus in 2, and influenza A (H3N2) in 1. The multiplex RT-PCR assay is recommended to clinical laboratories that are considering adoption of a molecular technique for viral diagnosis.


2021 ◽  
Vol 9 (1) ◽  
pp. 44-45
Author(s):  
Dinesh Kumar

Recently, an argument was put forth because a symptomatic and positive patient for CoVID-19 turned tested negative after 7 days, so discharged from the hospital. Both at the time of admission and discharge real-time reverse transcriptase Polymerase Chain Reaction (RT-PCR) was done for testing of CoVID-19. Immediately, patient again developed respiratory symptoms and was admitted to hospital again. Amidst of current CoVID-19 pandemic, a question was asked “What is the specificity of the Real Time-Polymerase Chain Reaction (RT-PCR) test for COVID-19?” with an assumption that what if at the time of discharge the disease is present in patient but test turned out to be negative? In response to that a counter statement was posed that “It is the sensitivity that should be asked rather than specificity”. It was based on the implication of primary question that was implying false negative report of the RT-PCR. It means, since patient was discharged with negative result that could be false negative.


2020 ◽  
Vol 66 (8) ◽  
pp. 1116-1121 ◽  
Author(s):  
Ertuğrul Güçlü ◽  
Mehmet Koroglu ◽  
Yusuf Yürümez ◽  
Hande Toptan ◽  
Elif Kose ◽  
...  

SUMMARY BACKGROUND Healthcare personnel are at risk of becoming infected while taking upper and/or lower respiratory tract specimens. Therefore, there is a need for sampling methods that do not risk infecting them. In this study, we aimed to compare the saliva and Oro-Nasopharyngeal Swab (ONS) sampling methods. METHODS Patients were divided into three groups. Group 1 included patients whose diagnosis of COVID-19 was confirmed by polymerase chain reaction (PCR). Group 2 included patients with COVID-19 compatible findings in lung computed tomography (CT), but with a negative PCR. Group 3 included patients who presented to the emergency department with COVID-19 compatible complaints but had normal CT. Saliva and ONS samples were taken on the third day of hospitalization in groups 1 and 2, whereas in group 3, they were taken at the time of admission to the hospital. RESULTS A total of 64 patients were included in the study. The average age was 51.04 ± 17.9 years, and 37 (57.8%) were male. SARS-CoV-2 was detected in 27 (42.2%) patients’ saliva samples. While the sensitivity and positive predictive value of saliva samples were 85.2%, specificity and negative predictive value were 89.2%. The value of kappa was in substantial agreement (0.744), and it was found statistically significant (<0.001). CONCLUSIONS Saliva samples can be used instead of ONS samples in detecting SARS-CoV-2. Investigating SARS-CoV-2 with saliva is cheaper, easier for the patient and overall, and, most importantly, it poses much less risk of SARS-CoV-2 contamination to healthcare personnel.


2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.


2020 ◽  
Author(s):  
Thomas Tschoellitsch ◽  
Martin Dünser ◽  
Carl Böck ◽  
Karin Schwarzbauer ◽  
Jens Meier

Abstract Objective The diagnosis of COVID-19 is based on the detection of SARS-CoV-2 in respiratory secretions, blood, or stool. Currently, reverse transcription polymerase chain reaction (RT-PCR) is the most commonly used method to test for SARS-CoV-2. Methods In this retrospective cohort analysis, we evaluated whether machine learning could exclude SARS-CoV-2 infection using routinely available laboratory values. A Random Forests algorithm with 1353 unique features was trained to predict the RT-PCR results. Results Out of 12,848 patients undergoing SARS-CoV-2 testing, routine blood tests were simultaneously performed in 1528 patients. The machine learning model could predict SARS-CoV-2 test results with an accuracy of 86% and an area under the receiver operating characteristic curve of 0.90. Conclusion Machine learning methods can reliably predict a negative SARS-CoV-2 RT-PCR test result using standard blood tests.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  

AbstractNowadays there is an ongoing acute respiratory outbreak caused by the novel highly contagious coronavirus (COVID-19). The diagnostic protocol is based on quantitative reverse-transcription polymerase chain reaction (RT-PCR) and chests CT scan, with uncertain accuracy. This meta-analysis study determines the diagnostic value of an initial chest CT scan in patients with COVID-19 infection in comparison with RT-PCR. Three main databases; PubMed (MEDLINE), Scopus, and EMBASE were systematically searched for all published literature from January 1st, 2019, to the 21st May 2020 with the keywords "COVID19 virus", "2019 novel coronavirus", "Wuhan coronavirus", "2019-nCoV", "X-Ray Computed Tomography", "Polymerase Chain Reaction", "Reverse Transcriptase PCR", and "PCR Reverse Transcriptase". All relevant case-series, cross-sectional, and cohort studies were selected. Data extraction and analysis were performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5. Among 1022 articles, 60 studies were eligible for totalizing 5744 patients. The overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan compared to RT-PCR were 87% (95% CI 85–90%), 46% (95% CI 29–63%), 69% (95% CI 56–72%), and 89% (95% CI 82–96%), respectively. It is important to rely on the repeated RT-PCR three times to give 99% accuracy, especially in negative samples. Regarding the overall diagnostic sensitivity of 87% for chest CT, the RT-PCR testing is essential and should be repeated to escape misdiagnosis.


Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal

<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.


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