scholarly journals Development of rapid immunochromatographic strip test for the detection of porcine epidemic diarrhoea virus

2017 ◽  
Vol 181 (22) ◽  
pp. 596-601 ◽  
Author(s):  
Kwang-Soo Lyoo ◽  
Minjoo Yeom ◽  
Jungho Kim ◽  
Donghyuk Kim ◽  
Gunwoo Ha ◽  
...  

Porcine epidemic diarrhoea virus (PEDV) causes acute and severe watery diarrhoea and dehydration, as well as 50–100 per cent mortality in piglets. For the PEDV diagnosis, a rapid test kit that is specific and sensitive to PEDV is critical to monitor this disease at pig farms. The present study aimed to develop an immunochromatographic assay (ICA) strip test for detecting PEDV in faecal swabs. The newly developed diagnostic test showed a detection limit of 104.0TCID50/ml of PEDV. Using faecal swab samples, the relative sensitivity and specificity of the ICA kit were 95.0 per cent and 98.6 per cent, respectively, compared with those of real-time RT-PCR. In samples from piglets experimentally infected with PEDV, the results showed 100 per cent agreement with those found by real-time RT-PCR. Our developed test strip will be useful for rapid diagnosis and can be used for epidemiological surveillance of PEDV infection.

2016 ◽  
Vol 60 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Yi-Xuan Hou ◽  
Chun Xie ◽  
Kang Wang ◽  
Yu-Ting Zhao ◽  
Yang-Yang Xie ◽  
...  

AbstractIntroduction:A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.Material and Methods:Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.Results:The developed quantitative PCR assay detected viral titres as low as 0.1 TCID50with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive.Conclusion:This approach is suitable for clinical sample identification and pathogenesis studies.


2017 ◽  
Vol 33 (1) ◽  
pp. 8-15
Author(s):  
LR Barman ◽  
RD Sarker ◽  
BC Das ◽  
EH Chowdhury ◽  
PM Das ◽  
...  

A virological survey for avian influenza (AI) and Newcastle disease (ND) was conducted in two selected live bird markets (LBMs), namely Kaptan Bazar and Karwan Bazar in Dhaka city, Bangladesh from August 2011 to July 2012. A total of 513 dead chickens were collected. An immune-chromatographic rapid antigen test for Type A influenza virus and both conventional and real time RT-PCR were used for the detection and characterization of AI and ND viruses. All carcasses were first screened by the rapid antigen test kit and 93 were positive for Type A influenza virus. RT-PCR on a representative number of rapid antigen test positive samples (n = 24) confirmed the presence of Type A influenza virus and mostly H5 influenza virus (22 out of 24 tested samples). Influenza rapid test negative samples (n = 420) were subjected to routine necropsy. Heat stress, suffocation and physical injury were the most common cause of mortality (163 cases), followed by ND, suspected to be the cause of 85 deaths. On molecular investigation of these 85 samples, the presence of ND virus was confirmed in 59 and AI virus in 6; 15 were negative for both ND and AI viruses and 5 were unsuitable for investigation. Among the 59 ND confirmed cases 18 also contained AI virus. In summary, out of 513 carcasses 117 (22.81%) contained AI virus and 59 (11.50%) contained ND virus. Eighteen (3.51%) carcasses contained both AI and ND viruses. The findings suggest that both AI and ND should be considered as major threats to the poultry industry.Bangl. vet. 2016. Vol. 33, No. 1, 8-15


2018 ◽  
Vol 19 (10) ◽  
pp. 3013 ◽  
Author(s):  
Trinh Thuy Tien ◽  
Hyun Park ◽  
Hien Tuong ◽  
Seung-Taek Yu ◽  
Du-Young Choi ◽  
...  

Human respiratory syncytial virus (RSV) is one of the most common viruses infecting the respiratory tracts of infants. The rapid and sensitive detection of RSV is important to minimize the incidence of infection. In this study, novel monoclonal antibodies (mAbs; B11A5 and E8A11) against RSV nucleoprotein (NP) were developed and applied to develop a rapid fluorescent immunochromatographic strip test (FICT), employing europium nanoparticles as the fluorescent material. For the FICT, the limits of detection of the antigen and virus were 1.25 µg/mL and 4.23 × 106 TCID50/mL, respectively, corresponding to 4.75 × 106 ± 5.8 ×105 (mean ± SD) RNA copy numbers per reaction mixture for RSV NP. A clinical study revealed a sensitivity of 90% (18/20) and specificity of 98.18% (108/110) for RSV detection when comparing the performance to that of reverse transcription polymerase chain reaction (RT-PCR), representing a 15% improvement in sensitivity over the SD Bioline rapid kit. This newly developed FICT could be a useful tool for the rapid diagnosis of RSV infection.


Author(s):  
Jesha Mundodan ◽  
Samina Hasnain ◽  
Hayat Khogali ◽  
Soha Shawqi Al Bayat ◽  
Dina Ali ◽  
...  

Background: In response to the growing coronavirus disease 2019 (COVID-19) pandemic and the shortage of laboratory based molecular testing capacity and reagents, multiple diagnostic test manufacturers have developed rapid and easy to use devices to facilitate testing outside laboratory settings. These kits are either based on detection of proteins from SARS-CoV-2 virus or detection of antigen or human antibodies generated in response to the infection. However, it is important to understand their performance characteristics and they must be validated in the local population setting.Design and Methods: The objective is to assess the validity of the rapid test for IgG and IgM immunoglobulins compared to the current gold standard reverse transcription polymerase chain reaction (RT-PCR) test. A total of 16951 asymptomatic individuals were tested by the Ministry of Public Health track-and-trace team using both rapid immunodiagnostic test and RT-PCR as part of screening across various random settings with potential risk of community interaction prior to gradual lifting of restrictions in Qatar.  Rapid test was considered to be posiive if both IgG and IgM are positive, while only IgG/IgM positive was considered as rapid test negative. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were calculated.Results: The sensitivity of rapid test kit was found to be 0.9%, whereas the specificity was found to be 97.8%. the PPV was found to be 0.3% whereas the NPV was found to be 99.4%.Conclusion: Based on the outcome and results of the study, it appears that the sensitivity and PPV of the rapid antibody test are low. As such, this test is not recommended for use to assist in taking clinic-based decisions or decisions related to quarantine/isolation.


2020 ◽  
Vol 54 ◽  
Author(s):  
Aldrich Ivan Lois D. Burog ◽  
Clarence Pio Rey C. Yacapin ◽  
Renee Rose O. Maglente ◽  
Anna Angelica Macalalad-Josue ◽  
Elenore Judy B. Uy ◽  
...  

KEY FINDINGS Current evidence does NOT support use of IgM/IgG rapid test kits for the definitive diagnosis of COVID-19 in currently symptomatic patients. • The present standard for diagnosis of COVID-19 is through qualitative detection of COVID-19 virus nucleic acid via reverse transcription polymerase chain reaction (RT-PCR). • Due to long turnaround times and complicated logistical operations, a rapid and simple field test alternative is needed to diagnose and screen patients. • An alternative to the direct detection and measurement of viral load (RT-PCR) is the qualitative detection of specific antibodies to COVID-19. ELISA (discussed in a separate rapid review) and lateral flow immunoassay (LFIA) IgM/IgG rapid test kits are two currently available, qualitative, antibody tests for COVID-19. • Two low quality clinical trials showed that there is insufficient evidence to support the use of IgM/IgG rapid test kits for the definitive diagnosis of COVID-19. Diagnostic accuracy varies greatly depending on the timing of the test. The test performed very poorly during the early phase of the disease (i.e., less than eight days from onset of symptoms). • Existing guidelines do not recommend serologic antibody tests for the diagnosis of COVID-19 in currently symptomatic patients.


2010 ◽  
Vol 88 (1) ◽  
pp. 166-168 ◽  
Author(s):  
Enrica Sozzi ◽  
Andrea Luppi ◽  
Davide Lelli ◽  
Ana Moreno Martin ◽  
Elena Canelli ◽  
...  

Author(s):  
Retno Handajani ◽  
Mochammad Thaha ◽  
Mochamad Amin ◽  
Citrawati Dyah Kencono Wungu ◽  
Edhi Rianto ◽  
...  

Anti-Human Immunodeficiency Virus (Anti-HIV) was performed from 100 plasma Chronic Kidney Disease (CKD) stage 5 patientswith continuous hemodialysis (HD) at the Hemodialysis Instalation Dr Soetomo hospital, Surabaya, Indonesia, using three (3) kind ofreagents: Tri-line HIV Rapid test Device from Acon for HIV 1/2/O as strips form, Foresight HIV 1/2/O Antibody EIA Test Kit from Aconand Anti-HIV 1+2/Subtype O ELISA from Axiom. HIV RNA and HIV subtype were detected by Reverse Transcription Polymerase ChainReaction (RT-PCR) based on HIV gag region and analysis of DNA result. Seventy three % patients were hemodialysed twice in a week andonly 14% with duration more than five (5) years. Most of the patients (43%) were hemodialysed between 100−300 times. From the 100plasma samples was obtained only one (1%) man patient plasma sample with positive anti-HIV. A weak positive of RT-PCR result wasnot succeed to be sequenced for determining the HIV subtype. This cause was suspected due to low levels of HIV RNA in blood. The resultsof this study was expected can be used as an additional management consideration of hemodialysis patients at the Hemodialysis Unit.


2020 ◽  
Vol 64 (1) ◽  
pp. 25-32 ◽  
Author(s):  
Yi-Ran Luo ◽  
Shu-Ting Zhou ◽  
Liang Yang ◽  
Yuan-Ping Liu ◽  
Sheng-Yao Jiang ◽  
...  

AbstractIntroductionPorcine epidemic diarrhoea virus (PEDV) infection causes watery diarrhoea, vomiting, anorexia, and weight loss, especially among neonatal piglets, inflicting on them morbidity and mortality potentially reaching 90%–100%. Despite it being known that certain mammalian cell phases are arrested by PEDV, the mechanisms have not been elucidated, and PEDV pathogenesis is poorly understood. This study determined the effect of an epidemic PEDV strain on cell cycle progression.Material and MethodsWe observed the effect of the PEDV SHpd/2012 strain on an infected Vero cell cycle through flow cytometry and Western blot, investigating the interrelationships of cell-cycle arrest, the DNA damage–signalling pathway caused by PEDV and the phosphorylation levels of the key molecules Chk.2 and H2A.X involved upstream and downstream in this pathway.ResultsPEDV induced Vero cell-cycle arrest at the G1/G0 phase. The phosphorylation levels of Chk.2 and H2A.X increased with the prolongation of PEDV infection, and no significant cell-cycle arrest was observed after treatment with ATM or Chk.2 inhibitors. The proliferation of PEDV was also inhibited by treatment with ATM or Chk.2 inhibitors.ConclusionPEDV-induced cell-cycle arrest is associated with activation of DNA damage–signalling pathways. Our findings elucidate the molecular basis of PEDV replication and provide evidence to support further evaluation of PEDV pathogenesis.


2020 ◽  
Author(s):  
Oluwafemi Ige ◽  
Ayuba Sunday Buru ◽  
Tanko Zainab Lamido ◽  
Tahir Mohammed ◽  
Livingstone Dogara ◽  
...  

AbstractThe emergence of the RNA virus SARS-CoV2, the causative agent of COVID-19 and its declaration by the World Health Organization (WHO) as a pandemic has disrupted the delicate balance in health indices globally. Its attendant immune dysregulation and pathobiology is still evolving. Currently, real time PCR is the gold standard diagnostic test, however there are several invalidated antibody-based tests available for possible community screening. With ongoing community transmission in Nigeria, neither the true burden of COVID-19 nor the performance of these kits is presently known. This study therefore, compared the performance of the SARS CoV2 antibody test and the real time Polymerase Chain Reaction (Rt-PCR) in the diagnosis of COVID-19. For the purpose of this evaluation, we used the diagnostic test kit by Innovita® Biological Technology CO., LTD China, a total of 521 venous blood samples were collected from consenting patients for the SARS COVID-19 rapid diagnostic kit and Oral and Nasopharyngeal swabs were collected and analyzed using the real time Polymerase chain reaction technique for nucleic acid detection and quantification.


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