Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates

Robardet, Emmanuelle; Andrieu, S; Rasmussen, Thomas Bruun; Dobrostana, M; Horton, Daniel L.; Hostnik, Peter; Jacevičienė, Ingrida; Juhász, Tamás; Müller, Thomas; Mutinelli, Franco; Servat, Alexandre; Smreczak, Marcin; Vanek, E.; Vázquez-Morón, Sonia; Cliquet, Florence

doi:10.1016/j.jviromet.2013.03.027

Cited by 20 publications

(17 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The questionnaire revealed several technical variations particularly in the application of the DFA test protocol. Although studies reporting that outcomes of the DFA test can be significantly affected by various factors such as the fluorescent conjugate [28], the cover glass mounting medium and its associated pH [44], and also, the microscope and type of filters in used [45], we did not observe any influence of technical steps for both DFA test and RT-PCR, very likely due the low number of discordant results. Information regarding the maintenance of equipment, the type of fluorescent microscope and filters in use were not collected during this PT.…”

Section: Discussioncontrasting

confidence: 67%

An inter-laboratory trial as a tool to increase rabies diagnostic capabilities of Sub-Saharan African Veterinary laboratories

et al. 2020

View full text Add to dashboard Cite

To achieve the goal of eliminating dog-mediated human rabies deaths by 2030, many African countries have agreed to list rabies as a priority zoonotic disease and to undertake both short and long-term control programs. Within this context, reliable local diagnosis is essential for the success of field surveillance systems. However, a harmonized, sustainable and supportive diagnostic offer has yet to be achieved in the continent. We herewith describe the organization and outcome of a proficiency test (PT) for the post-mortem diagnosis of rabies in animals, involving thirteen veterinary laboratories and one public health laboratory in Africa. Participants were invited to assess both the performance of the Direct Fluorescent Antibody (DFA) test and of a conventional RT-PCR. From the submitted results, while thirteen laboratories proved to be able to test the samples through DFA test, eleven performed the RT-PCR method; ten applied both techniques. Of note, the number of laboratories able to apply rabies RT-PCR had increased from four to ten after the exercise. Importantly, results showed a higher proficiency in applying the molecular test compared to the DFA test (concordance, sensitivity and specificity: 98.2%, 96.97% and 100% for RT-PCR; 87.69%, 89.23% and 86.15% for DFA test), indicating the feasibility of molecular methods to diagnose animal pathogens in Africa. Another positive outcome of this approach was that negative and positive controls were made available for further in-house validation of new techniques; in addition, a detailed questionnaire was provided to collect useful and relevant information on the diagnostic procedures and biosafety measures applied at laboratory level.

show abstract

Section: Discussioncontrasting

confidence: 67%

An inter-laboratory trial as a tool to increase rabies diagnostic capabilities of Sub-Saharan African Veterinary laboratories

et al. 2020

View full text Add to dashboard Cite

show abstract

“…[40] and S h a r m a et al [33], gave 17 % and 64.7 % positive results respectively by RIDT on fresh samples. Generally, the accuracy of rabies diagnosis is dependent on the quality of the sample [13,6], the type of anti-rabies conjugate used [31], virus antigen distributions in the brain and areas of the brain tested [4].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Enzyme Linked Immuno-Sorbent Assay and Rapid Immuno-Diagnostic Test for Rabies Antigen Detection in Archived Dog Brain Tissues

Okoh

Kazeem²,

Kia

et al. 2018

Folia Veterinaria

View full text Add to dashboard Cite

Rabies urgently requires strengthening of new and existing diagnostic methodology in order to overcome the threat it poses. We evaluated the Enzyme Linked Immuno-Sorbent Assay (ELISA) and the Rapid Immunodiagnostic Test (RIDT) in detecting rabies viral antigens, comparing both tests with the Direct Fluorescent Antibody Test (DFAT) which is the gold standard in rabies diagnosis. Fifty dog brain tissues collected from the archives of the Central Diagnostic Laboratory, National Veterinary Research Institute, Vom, Nigeria, were utilized for this study. ELISA performed better than RIDT and recorded equivalent result with DFAT as compared with RIDT. There was a 96 % agreement between ELISA and DFAT for rabies antigen detection (concordance coefficient 78 % : 95 % C. I. 0.6366 to 0.8654) while there was a 54 % agreement between RIDT and DFAT (concordance coefficient 17 % : 95 % C. I. 0.05138—0.2752). Compared to DFAT, the sensitivities of ELISA and RIDT were 95.5 % and 47.6 %, respectively, and the specificities of ELISA and RIDT were 100 % and 87.5 % respectively. The simple Cohen’s kappa coefficient for ELISA related to the DFAT was found to be 0.834 (95 % C. I. 0.613—1.0). For RIDT, the Kappa value was 0.170 (95 % C. I. 0.003—0.337). The ELISA is as reliable a diagnostic method as the DFAT which is the gold standard for rabies diagnosis. It has an advantage of being able to analyse large number of samples at the same time, making it more suitable for epidemiological studies and for laboratories that cannot perform the DFAT. The unsatisfactory result of RIDT in this study reiterates the need to perform an adequate test validation before it can be used in the laboratory for rabies diagnosis.

show abstract

“…While no significant associations were seen between differences in survey responses and laboratory performance, this may be due to the small sample size, as even small methodological changes have been shown to affect the sensitivity of the DFA test including the mounting medium [16–17], laboratorian’s expertise level, number of employees reading the slides, the anti-rabies conjugate utilized, specificity controls used and working performance of fluorescence microscope [18, 19]. …”

Section: Discussionmentioning

confidence: 99%

“…For the current exercise, laboratories used commercial (65%) or in-house (35%) conjugates. A study of 12 rabies reference laboratories in Europe demonstrated that the variability of conjugates could potentially lead to discordant results and influence assay sensitivity [19]. The OIE manual states that the conjugates should be fully validated for specificity and sensitivity before use, including their ability to detect lyssaviruses other than rabies [3]; it is unclear whether such validation has been completed for the in-house conjugates used in the LAC region.…”

Section: Discussionmentioning

confidence: 99%

An inter- laboratory proficiency testing exercise for rabies diagnosis in Latin America and the Caribbean

et al. 2017

View full text Add to dashboard Cite

The direct fluorescent antibody test (DFA), is performed in all rabies reference laboratories across Latin America and the Caribbean (LAC). Despite DFA being a critical capacity in the control of rabies, there is not a standardized protocol in the region. We describe the results of the first inter-laboratory proficiency exercise of national rabies laboratories in LAC countries as part of the regional efforts towards dog-maintained rabies elimination in the American region. Twenty three laboratories affiliated to the Ministries of Health and Ministries of Agriculture participated in this exercise. In addition, the laboratories completed an online questionnaire to assess laboratory practices. Answers to the online questionnaire indicated large variability in the laboratories throughput, equipment used, protocols availability, quality control standards and biosafety requirements. Our results will inform actions to improve and harmonize laboratory rabies capacities across LAC in support for the regional efforts towards elimination of dog-maintained rabies.

show abstract

Comparative assay of fluorescent antibody test results among twelve European National Reference Laboratories using various anti-rabies conjugates

Cited by 20 publications

References 10 publications

An inter-laboratory trial as a tool to increase rabies diagnostic capabilities of Sub-Saharan African Veterinary laboratories

An inter-laboratory trial as a tool to increase rabies diagnostic capabilities of Sub-Saharan African Veterinary laboratories

Evaluation of Enzyme Linked Immuno-Sorbent Assay and Rapid Immuno-Diagnostic Test for Rabies Antigen Detection in Archived Dog Brain Tissues

An inter- laboratory proficiency testing exercise for rabies diagnosis in Latin America and the Caribbean

Contact Info

Product

Resources

About