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 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 Comparison of PanBio Dengue Duo Enzyme-Linked Immunosorbent Assay (ELISA) and MRL Dengue Fever Virus Immunoglobulin M Capture ELISA for Diagnosis of Dengue Virus Infections in Southeast Asia

1999 ◽  
Vol 6 (5) ◽  
pp. 705-712 ◽  
Author(s):  
Andrea J. Cuzzubbo ◽  
David W. Vaughn ◽  
Ananda Nisalak ◽  
Tom Solomon ◽  
Siripen Kalayanarooj ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Laboratory Diagnosis ◽  
Immunoglobulin M ◽  
Fever Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Capture Elisa

ABSTRACT The performances of the MRL dengue fever virus immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) and the PanBio Dengue Duo IgM capture and IgG capture ELISA were compared. Eighty sera from patients with dengue virus infections, 24 sera from patients with Japanese encephalitis (JE), and 78 sera from patients with nonflavivirus infections, such as malaria, typhoid, leptospirosis, and scrub typhus, were used. The MRL test showed superior sensitivity for dengue virus infections (94 versus 89%), while the PanBio test showed superior specificity for JE (79 versus 25%) and other infections (100 versus 91%). The PanBio ELISA showed better overall performance, as assessed by the sum of sensitivity and specificity (F value). When dengue virus and nonflavivirus infections were compared, F values of 189 and 185 were obtained for the PanBio and MRL tests, respectively, while when dengue virus infections and JE were compared, F values of 168 and 119 were obtained. The results obtained with individual sera in the PanBio and MRL IgM ELISAs showed good correlation, but this analysis revealed that the cutoff value of the MRL test was set well below that of the PanBio test. Comparing the sensitivity and specificity of the tests at different cutoff values (receiver-operator analysis) revealed that the MRL and PanBio IgM ELISAs performed similarly in distinguishing dengue virus from nonflavivirus infections, although the PanBio IgM ELISA showed significantly better distinction between dengue virus infections and JE. The implications of these findings for the laboratory diagnosis of dengue are discussed.

scholarly journals Epidemiological Survey and Screening Strategy for Dengue Virus in Blood Donors from Yunnan Province

2020 ◽  
Author(s):  
LING LI ◽  
YING LI ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Prevalence Rate ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Igm Antibodies ◽  
Blood Center ◽  
Nonstructural Protein 1 ◽  
Elisa Testing

Abstract BACKGROUND Dengue virus (DENV) can be transmitted through blood transfusion. DENV was not screened regularly in Xishuangbanna Blood Center. This study was conducted in Xishuangbanna Blood Center with an attempt to develop DENV screening strategies in one of China’s high-incidence areas.METHODS Blood samples were collected randomly between June 2019 and August 2019. These samples were first screened for dengue IgG and IgM antibodies using enzyme-linked immunosorbent assay (ELISA). All reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real time polymerase-chain-reaction (RT-PCR) assay. After RT-PCR assay, these samples were further tested for soluble nonstructural protein 1 (NS1) using colloidal gold method. The demographic data of DENV positive donors were collected.RESULTS A total of 2,254 donor samples were collected and tested for dengue IgG and IgM antibodies by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-IgG and/or anti-IgM reactive, with a serological prevalence rate of 26.53%. Among all the donor samples, 26 were RT-PCR positive and/or NS1 positive. Moreover, there were significant differences in the prevalence rate of DENV in terms of occupation (P=0.001), education(P<0.001) and ethnicity (P=0.026). CONCLUSION The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides the first-hand data about the prevalence of DENV and allows development of a screening strategy for clinical use.

scholarly journals Evaluation of a Novel Reporter Virus Neutralization Test for Serological Diagnosis of Zika and Dengue Virus Infection

2017 ◽  
Vol 55 (10) ◽  
pp. 3028-3036 ◽  
Author(s):  
Chao Shan ◽  
Daniel A. Ortiz ◽  
Yujiao Yang ◽  
Susan J. Wong ◽  
Laura D. Kramer ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Neutralizing Antibodies ◽  
Neutralization Test ◽  
Reference Method ◽  
Laboratory Diagnosis ◽  
Virus Neutralization Test ◽  
Virus Neutralization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reporter Virus ◽  
Screening Assay

ABSTRACT Currently, the laboratory diagnosis of Zika virus (ZIKV) infection is primarily through the detection of ZIKV RNA or antibodies against ZIKV proteins. The detection of viral RNA is highly sensitive and specific, but periods of viremia and viruria are brief, limiting the utility of ZIKV RNA assays. Instead, most ZIKV infections are diagnosed serologically, using an IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for screening, followed by a confirmatory plaque reduction neutralization test (PRNT). Typical turnaround times vary, due to assay incubation periods and a lack of clinical laboratories performing these tests. Recently, a novel luciferase-ZIKV- and -dengue virus (DENV)-based serological assay, which considerably improves the turnaround times and throughput for ZIKV diagnosis, was described. Using the traditional PRNT as a reference method, we evaluated the performance characteristics of the reporter virus neutralization test (RVNT) with 258 clinical serum specimens. The ZIKV RVNT produced primary ZIKV screening and secondary confirmation results in 4 days, with 100% reproducibility. As a screening assay, the ZIKV RVNT displayed excellent diagnostic accuracy, sensitivity, and specificity of 98.2%, 100%, and 98.1%, respectively. As a confirmatory assay, the ZIKV RVNT titers displayed 93.1% agreement with the traditional ZIKV PRNT titers. Overall, the RVNT accurately and reliably detects neutralizing antibodies in patient serum specimens, with improved turnaround times, and can be used for the serological detection of ZIKV infections. Due to the homogeneous 96-well format, the RVNT has also significantly improved the assay throughput to allow testing of a large number of specimens in a single run.

scholarly journals Performance Evaluation of Commercial Dengue Diagnostic Tests for Early Detection of Dengue in Clinical Samples

2017 ◽  
Vol 2017 ◽  
pp. 1-4 ◽  
Author(s):  
Tuan Nur Akmalina Mat Jusoh ◽  
Rafidah Hanim Shueb
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Dengue Infection ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
Kappa Agreement

The shattering rise in dengue virus infections globally has created a need for an accurate and validated rapid diagnostic test for this virus. Rapid diagnostic test (RDT) and reverse transcription-polymerase chain reaction (RT-PCR) diagnostic detection are useful tools for diagnosis of early dengue infection. We prospectively evaluated the diagnostic performance of nonstructural 1 (NS1) RDT and real-time RT-PCR diagnostic kits in 86 patient serum samples. Thirty-six samples were positive for dengue NS1 antigen while the remaining 50 were negative when tested with enzyme-linked immunosorbent assay (ELISA). Commercially available RDTs for NS1 detection, RTK ProDetect™, and SD Bioline showed high sensitivity of 94% and 89%, respectively, compared with ELISA. GenoAmp® Trioplex Real-Time RT-PCR and RealStar® Dengue RT-PCR tests presented a comparable kappa agreement with 0.722. The result obtained from GenoAmp® Real-Time RT-PCR Dengue test showed that 14 samples harbored dengue virus type 1 (DENV-1), 8 samples harbored DENV-2, 2 samples harbored DENV-3, and 1 sample harbored DENV-4. 1 sample had a double infection with DENV-1 and DENV-2. The NS1 RDTs and real-time RT-PCR tests were found to be a useful diagnostic for early and rapid diagnosis of acute dengue and an excellent surveillance tool in our battle against dengue.

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.

scholarly journals Enzyme linked immunosorbent assay (ELISA) for the detection of IgG, IgM, IgE and IgA against Cysticercus cellulosae in cerebrospinal fluid of patients with neurocysticercosis

2002 ◽  
Vol 60 (2B) ◽  
pp. 400-405 ◽  
Author(s):  
Newton Satoru Odashima ◽  
Osvaldo Massaiti Takayanagui ◽  
José Fernando de Castro Figueiredo
Keyword(s):  
Cerebrospinal Fluid ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Neurological Diseases ◽  
Active Form ◽  
Clinical Manifestations ◽  
Enzyme Linked Immunosorbent Assay ◽  
Inactive Form ◽  
Cysticercus Cellulosae ◽  
Inflammation Signs

The objective of this study was to analyze different immunoglobulins classes (IgG, IgM, IgE and IgA) against Cysticercus cellulosae in the cerebrospinal fluid (CSF), through enzyme linked immunosorbent assay (ELISA), correlating them to clinical and tomographic profiles in patients with neurocysticercosis (NCC). Eighty-five specimens of CSF were obtained from 43 cases with NCC (26 with the active form and 17 with the inactive form) and from 42 patients with other neurological diseases. The inactive form of NCC presented a profile in CSF similar to the group without NCC. The active form of NCC presented elevation of specific immunoglobulins (IgG, IgM, IgE, and IgA) in decreasing order, with the highest values being detected among the cases with intraventricular cysts, or with inflammation signs in CSF or in those with multiple clinical manifestations. The highest sensitivity and specificity were obtained with ELISA-IgG (88.5% and 93.2%, respectively). This study confirmed the importance of ELISA in the immunologic diagnosis of NCC.

scholarly journals Validation of Selected Commercial Serological Assays for Diagnosis of COVID-19

2020 ◽  
Author(s):  
Salma Younes ◽  
Hadeel Al-Jighefee ◽  
Farah Shurrab ◽  
Duaa Al-Sadeq ◽  
Hadi Yassine ◽  
...  
Keyword(s):  
Neutralization Test ◽  
Cross Reactivity ◽  
Clinical Settings ◽  
Igg Antibodies ◽  
Excellent Correlation ◽  
Rt Pcr ◽  
Rapid Assays ◽  
Igm Elisa ◽  
Rapid Tests ◽  
Antibody Tests

As researchers around the globe rush to put the available antibody tests to use, concerns have been raised about their precision. This study aimed to evaluate and compare the performance of selected commercial & automated serological assays that are widely used in different clinical settings in Qatar. We validated the performance of five commercial IgG and IgM ELISA kits, three fully automated immunoassays, and two commercial rapid tests. The sensitivity of all assays was compared to RT-PCR and a surrogate virus neutralization test (sVNT). In addition, cross-reactivity was investigated. Among the evaluated kits, Lionex IgG assay demonstrated the best performance (~88% sensitivity and ~99 specificity). All automated assays showed an excellent correlation with the neutralization test with an overall agreement of 93.6-98.5%. The rapid assays demonstrated a very good performance in detecting IgG antibodies (86.0-88.0% sensitivity and 98.0-100% specificity).

scholarly journals Comparison of Four Serological Methods and Two Reverse Transcription-PCR Assays for Diagnosis and Surveillance of Zika Virus Infection

2018 ◽  
Vol 56 (3) ◽  
Author(s):  
Angel Balmaseda ◽  
José Victor Zambrana ◽  
Damaris Collado ◽  
Nadezna García ◽  
Saira Saborío ◽  
...  
Keyword(s):  
Reverse Transcription ◽  
Zika Virus ◽  
Clinical Manifestations ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Emerging Infections ◽  
Rt Pcr ◽  
Convalescent Phase ◽  
Reverse Transcription Pcr ◽  
Pcr Assays

ABSTRACTZika virus (ZIKV) is a mosquito-borne flavivirus that is responsible for recent explosive epidemics in the Americas. Notably, ZIKV infection during pregnancy has been found to cause congenital birth defects, including microcephaly, and ZIKV has been associated with Guillain-Barré syndrome in adults. Diagnosis and surveillance of Zika in the Americas have been challenging due to similar clinical manifestations and extensive antibody cross-reactivity with endemic flaviviral diseases, such as dengue. We evaluated four serological and two reverse transcription-PCR (RT-PCR) methods in acute-phase (mean day, 1.8), early-convalescent-phase (mean day, 16.7), and late-convalescent-phase (mean, ~7 months) samples from the same individuals in a long-term pediatric cohort study in Nicaragua. Well-characterized samples from 301 cases of Zika, dengue, or non-Zika, nondengue febrile illnesses were tested. Compared to a composite reference, an in-house IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and the NIAID-Biodefense and Emerging Infections (BEI) MAC-ELISA measuring IgM yielded sensitivities of 94.5% and 70.1% and specificities of 85.6% and 82.8%, respectively. The NS1 blockade-of-binding ELISA measuring anti-ZIKV NS1 antibody levels yielded sensitivities of 85.0% and 96.5% and specificities of 91.4% and 92.6% at early and late convalescence, respectively. An inhibition ELISA detecting total anti-ZIKV antibodies had sensitivity and specificity values of 68.3% and 58.3% for diagnosis and 94.0% and 98.6% for measuring annual infection incidence. Finally, the ZCD and Trioplex real-time RT-PCR assays detecting Zika, chikungunya, and dengue viruses both yielded a sensitivity of 96.1% and specificity of 100%. Together, these assays resolve the urgent need for diagnostic and surveillance tools for countries affected by Zika virus infections.

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