scholarly journals Performance of Latex agglutination, ELISA and RT-PCR for diagnosis of Rotavirus infection

2018 ◽  
Vol 90 (2) ◽  
Author(s):  
Hosna Hamzavi ◽  
Azarakhsh Azaran ◽  
Manoochehr Makvandi ◽  
Sahar Karami ◽  
Mohammad Roayaei Ardakani ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Acute Gastroenteritis ◽  
Laboratory Diagnosis ◽  
Latex Agglutination ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Rotavirus Infection ◽  
Polymerase Chain ◽  
Major Factors ◽  
Positive Results

The rotavirus is one of the major factors of inducing the acute gastroenteritis infection in children under 5 years of age. The laboratory diagnosis is progress and bringing it under control as well as avoiding its diffusion. The purpose of the present study was to determine the performance of enzyme linked immunosorbent assay (ELISA) and Latex agglutination (LA) tests against reverse transcription-polymerase chain reaction (RT-PCR) for evaluating the children’s acute gastroenteritis by rotavirus. One hundred feces specimens were collected from February to May 2014 and analyzed by LA, ELISA and RT-PCR. In this study, the positive results for rotavirus detected by ELISA, LA and RT-PCR were 37, 43 and 27%, respectively. In addition, the result showed that the sensitivity and specificity of ELISA and LA were 74 and 85%, respectively, when compared to RT-PCR. For laboratory detection of Rotavirus infection, RT-PCR has the highest sensitivity and specificity but because of the high costs, ELISA and LA based kits with good performance, as shown by this study, can be preferred for the routine use.

scholarly journals Serological and Molecular Detection of Hepatitis B virus among patients referred to Kurdistan Center for Hepatology and Gastroenterology in Sulaimani City/Kurdistan Region of Iraq

2020 ◽  
Vol 4 (2) ◽  
pp. 117
Author(s):  
Raz Sirwan Abdulla ◽  
Salih Ahmed Hama
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hbv Dna ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
B Virus ◽  
Polymerase Chain ◽  
Pcr Techniques ◽  
Positive Results

Hepatitis B virus infection is caused by the hepatitis B virus, a major global health problem. This infection can lead to chronic conditions, followed by cirrhosis and hepatocellular carcinoma (HCC). The current study was aimed to detect HBV using serological and molecular techniques. During 2019, 300 blood samples were collected from Kurdistan Center for Hepatology and Gastroenterology in Sulaimani city. Enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) techniques were used for the detection of HBsAg and HBV DNA, respectively. Obtained results were revealed that 92 out of 300 tested patients (30.66%) seropositive for HBsAg. Among 92 seropositive patients, 53 were shown positive results for HBV DNA by RT-PCR. Dental clinic visiting and dialysis were among the important risk factors for HBV transmission. The vast majority of positive results were among males. Smokers showed relatively high rates of positive results. One-third of the referred patients who had liver complaints were positive for HBsAg. More than half of the seropositive patients showed RT-PCR positive results. It was concluded that the molecular method (RT-PCR) is more sensitive and gives a more accurate result than serology (ELISA). Therefore, it can be used as a diagnostic tool for HBV detection.

Investigation of Rotavirus infection in sheep usingserological and molecular techniques

2016 ◽  
Author(s):  
Volkan Yilmaz. ◽  
M.Ozkan Timurkan ◽  
Nuvit Coskun ◽  
Yakup Yildirim
Keyword(s):  
Molecular Detection ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Family Farms ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Rotavirus Infection ◽  
Scale Family ◽  
Polymerase Chain

In this study, serological and molecular research was conducted on the Rotavirus infection in domestic breeds of sheep at 2–3 years of age. The sheep included in the study were raised on small scale family units of less than 20 sheep per unit, in central Kars province and its districts (Susuz, Arpaçay, Kagizman and Selim) in the Northeast Anatolia region of Turkey. The blood and fecal samples were collected randomly from 450 sheep. They were analyzed for the presence of Rotavirus and the antibody against the virus using enzyme-linked immunosorbent assay (ELISA). The highest seropositive ratio (73.46%) was found in central Kars province. The seroprevalence of Rotavirus in sheep raised in the Kars region was determined to be 55.33%. Rotavirus was not detected in fecal samples with ELISA. Molecular detection of Rotavirus from fecal samples was done by reverse transcription polymerase chain reaction (RT-PCR) technique using specific generic primers for VP6 protein. Rotavirus could not be detected in RT-PCR. The data that were obtained showed that the infection spreads on small scale family farms. Based on this information, recommendations were made for controlling Rotavirus infection.

scholarly journals Challenges and Opportunities of Laboratory diagnosis of Dengue virus infection: a review

2019 ◽  
Author(s):  
Gaspary Oigen Mwanyika ◽  
Gerald Misinzo ◽  
Sima Rugarabamu ◽  
Calvin Sindato ◽  
Leonard Mboera
Keyword(s):  
Dengue Virus ◽  
Sensitivity And Specificity ◽  
Diagnostic Tests ◽  
New Technologies ◽  
Clinical Manifestations ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Igm Elisa ◽  
Rapid Tests

Abstract Background: Globally, dengue is one of the most important mosquito-borne viral diseases. Lack of effective vaccines and specific therapy against the disease threaten global health. Reliance on clinical diagnosis is complex due to clinical manifestations which resemble other diseases. This review examined various challenges of current dengue laboratory diagnoses, emerging technological opportunities and highlights considerations for future dengue diagnoses. Methods: A literature search from PubMed, Web of Science and Google Scholar databases was done from October 2018 to January 2019. Thematic descriptive analysis was done for all qualitative data and quantitative data analysis for computation of sensitivity and specificity of selected diagnostic tests at 95% confidence was done using R software (v3.4.4, mada package). The results: A total of 128 articles was reviewed. The current dengue laboratory diagnoses include (i) virus isolation (ii) detection of nucleic acid (iii) detection of non-structural protein 1(NS1) antigen and (iv) detection of anti-dengue antibodies. Assessment of diagnostic performance shows that reverse transcription-polymerase chain reaction (RT-PCR) and IgM antibody capture enzyme-linked immunosorbent assay (IgM ELISA) have high and consistent sensitivity (82.6% to 99.2% and 92.8% to 97.8%, respectively) and specificity (78.8% to 100% and 80.3% to 99%, respectively) compared to NS1 ELISA and NS1 commercial rapid tests with sensitivity (53.7% to 96.2% and 49.7% to 99.5%, respectively) and specificity (34.5% to 93.8% and 63.8% to 98.6%, respectively). Major challenges of dengue laboratory diagnosis include lack of reliable tests for routine purposes. Routine dengue tests are mainly serological, which are not suitable for discriminating dengue virus from other infecting flaviviruses due to cross-reactivity, narrow window of diagnosis due to short virus life cycle and inconsistent performance of commercial rapid tests. New technologies such as biosensors and nanobodies are being developed to improve sensitivity, specificity, detection time and reduce the cost. However, the performance of these new tools under field condition is unknown. Conclusion: Currently, RT-PCR and IgM ELISA are the most sensitive and specific dengue diagnostic tests, despite their limitations. Future research should explore emerging technologies to improve the sensitivity and specificity of dengue diagnostics.

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 Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  
Keyword(s):  
Real Time ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Current Season ◽  
Seed Market ◽  
Polymerase Chain ◽  
Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

scholarly journals Epidemiology and Control of Congo Fever in Sacrificial Animals of Pakistan

2019 ◽  
Vol 1 (2) ◽  
Author(s):  
Hafiz Muhammad Rizwan ◽  
Muhammad Sohail Sajid ◽  
Haider Abbas ◽  
Muhammad Fiaz Qamar ◽  
Qaiser Akram
Keyword(s):  
Tick Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Many Body ◽  
Rt Pcr ◽  
Body Tissues ◽  
Crimean Congo Haemorrhagic Fever ◽  
Polymerase Chain ◽  
And Control ◽  
Successful Prevention ◽  
Venereal Transmission

The cases and deaths due to Crimean-Congo haemorrhagic fever (CCHF) [49] virus commonly known as Congo virus (fatality rate 15%) have been reported throughout Pakistan from the last five years especially during religious occasion, Eid-ul-Azha. The annual increase in death rates due to CCHF demonstrate the importance of awareness of Congo fever at academia as well as public level. The symptoms of Congo fever which appear one to nine days after tick bite, include sudden high fever, muscle aches, abdominal pain, headache, dizziness, sore eyes, jaundice, mood swings, confusion, aggression, and sensitivity to light. The other signs include sore throat, joint pain, vomiting, diarrhea, hemorrhages, and bleeding from skin and large intestine. The Infection has been reported in many species of wild as well as domestic animals including hares, cattle, sheep, goats, dogs, mice and hedgehogs. At least 31 species of Hyalomma, Boophilus, Rhipicephalus, Dermacentor (Ixodidae: hard ticks) act as vector of CCHF in which transovarial, transstadial and venereal transmission occurs. The virus attacks the immune system of the host and influences the immune cells. The Congo fever virus can be isolated from blood, plasma and many body tissues (kidneys, liver, spleen, lungs, brain and bone marrow). Mice inoculation, enzyme-linked immunosorbent assay (ELISA), reverse transcription polymerase chain reaction (RT-PCR) can be used for detection of the infection. Furthermore, IgM and IgG antibodies against CCHFV can also be detected and quantified. Education of general public, tick control with acaricides, use of anti-CCHFV immunoglobulin, usage of approved repellents to prevent tick bites, wearing neutral-coloured garments, application of a permethrin spray to the clothing, avoiding tall grasses and shrubs, applying sunscreen, avoiding direct contact with the blood or tissues of animals are the factors for successful prevention of the infection.

scholarly journals Diagnosis of Visceral Leishmaniasis by Enzyme-Linked Immunosorbent Assay Using Urine Samples

2002 ◽  
Vol 9 (4) ◽  
pp. 789-794 ◽  
Author(s):  
Mohammad Zahidul Islam ◽  
Makoto Itoh ◽  
S. M. Shamsuzzaman ◽  
Rusella Mirza ◽  
Farzana Matin ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Laboratory Diagnosis ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Serum Samples ◽  
Crude Antigen

ABSTRACT A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone-treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis.

Clinical Relevance of Annual Screening Using a Commercial Enzyme-Linked Immunosorbent Assay (SNAP 3Dx) for Canine Ehrlichiosis

2009 ◽  
Vol 45 (3) ◽  
pp. 118-124 ◽  
Author(s):  
Barbara C. Hegarty ◽  
Pedro Paulo Vissotto de Paiva Diniz ◽  
Julie M. Bradley ◽  
Leif Lorentzen ◽  
Edward Breitschwerdt
Keyword(s):  
Clinical Relevance ◽  
Deoxyribonucleic Acid ◽  
Immunofluorescence Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Active Infection ◽  
Chain Reaction ◽  
Canine Ehrlichiosis ◽  
Annual Screening ◽  
Polymerase Chain ◽  
Positive Results

Eighty-six dogs were selected based upon Ehrlichia (E.) canis SNAP 3Dx positive results to determine clinical relevance of annual E. canis screening. Immunofluorescence assay showed 72 (84%) of 86 dogs were seroreactive for E. canis. Polymerase chain reaction (PCR) revealed that 12 (14%) of 86 dogs had Ehrlichia deoxyribonucleic acid; seven had E. canis, four had E. ewingii, and one was coinfected with E. chaffeensis and E. ewingii. Thrombocytopenia (&lt;164,000 platelets/μL) was found in 28 (39%) of 72 dogs. In this study, thrombocytopenia was frequently detected in healthy Ehrlichia SNAP 3Dx-positive dogs, whereas active infection was infrequently confirmed by PCR. Therefore, treatment based upon screening results alone is not recommended.

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