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

Gourlaouen, Morgane; Angot, Angélique; Mancin, Marzia; Bebay, Charles; Soumare, Baba; Ellero, Francesca; Zecchin, Barbara; Leopardi, Stefania; Battisti, Cristian De; Terregino, Calogero; Benedictis, Paola De

doi:10.1371/journal.pntd.0008010

Cited by 8 publications

(8 citation statements)

References 33 publications

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“…Although dFAT is regarded as the gold standard method for rabies diagnosis in humans and animals [33,48], the results are sometimes affected by the examiner's experience and knowledge [33]. Furthermore, differences in processing methods or in the conjugate also influence the sensitivity and specificity of the test [13,15,49]. To maintain the quality and performance of dFAT in each laboratory, it is absolutely necessary to ensure accurate, reliable, and sustainable results via regular training by a reference laboratory.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the diagnostic accuracy of lateral flow devices as a tool to diagnose rabies in post-mortem animals

et al. 2020

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Implementation of lateral flow devices (LFDs) for rabies antigen detection is expected to improve surveillance through the efficient detection of rabid animals in resource-limited settings; however, the use of LFDs for diagnosis remains controversial because some commercially available kits show low sensitivity. Therefore, we compared the diagnostic efficacy of three LFDs (ADTEC, Bionote, and Elabscience kits) paralleled with the direct fluorescent antibody test (dFAT) using fresh samples and investigated the diagnostic accuracies. To do so, we evaluated rabies-suspected samples submitted to the Regional Animal Disease Diagnostic Laboratory III, Philippines. Furthermore, we conducted real-time RT-PCR and sequencing to measure the accuracy of field laboratory diagnosis. The total number of animals submitted during this study period was 184 cases, including negative control samples. Of these, 53.9% (84 cases) were positive in the dFAT. Dogs were the most common rabies-suspected animal (n = 135). The sensitivities of the ADTEC and Bionote kits were 0.88 (74 cases) and 0.95 (80 cases), respectively. The specificity of both kits was 1.00 (100 cases). Furthermore, the sensitivity and specificity of the ADTEC kit after directly homogenizing the samples in assay buffer without dilution in phosphate-buffered saline (ADTEC kit DM) were 0.94 (79 cases) and 1.00 (100 cases), respectively. By contrast, there were no positive results using the Elabscience kit among all dFAT-positive samples. The sensitivity and specificity of LFDs make these tests highly feasible if properly used. Therefore, LFD tests can be used to strengthen the surveillance of rabies-infected animals in endemic and resource-limited settings.

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Section: Discussionmentioning

confidence: 99%

Evaluation of the diagnostic accuracy of lateral flow devices as a tool to diagnose rabies in post-mortem animals

et al. 2020

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“…Often overlooked, rabies diagnosis has undergone a seeming renaissance of late 2 , 7 , 51 – 53 . Infection is confirmed by the finding of viral antigens, antibodies, amplicons, nucleic acids, or biomarkers in subjects with signs compatible with an encephalitis.…”

Section: Diagnostic Applicationsmentioning

confidence: 99%

Update on lyssaviruses and rabies: will past progress play as prologue in the near term towards future elimination?

Rohde

Rupprecht²

2020

Fac Rev

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Rabies is an ancient, much-feared, and neglected infectious disease. Caused by pathogens in the family Rhabdoviridae, genus Lyssavirus, and distributed globally, this viral zoonosis results in tens of thousands of human fatalities and millions of exposures annually. All mammals are believed susceptible, but only certain taxa act as reservoirs. Dependence upon direct routing to, replication within, and passage from the central nervous system serves as a basic viral strategy for perpetuation. By a combination of stealth and subversion, lyssaviruses are quintessential neurotropic agents and cause an acute, progressive encephalitis. No treatment exists, so prevention is the key. Although not a disease considered for eradication, something of a modern rebirth has been occurring within the field as of late with regard to detection, prevention, and management as well as applied research. For example, within the past decade, new lyssaviruses have been characterized; sensitive and specific diagnostics have been optimized; pure, potent, safe, and efficacious human biologics have improved human prophylaxis; regional efforts have controlled canine rabies by mass immunization; wildlife rabies has been controlled by oral rabies vaccination over large geographic areas in Europe and North America; and debate has resumed over the controversial topic of therapy. Based upon such progress to date, there are certain expectations for the next 10 years. These include pathogen discovery, to uncover additional lyssaviruses in the Old World; laboratory-based surveillance enhancement by simplified, rapid testing; anti-viral drug appearance, based upon an improved appreciation of viral pathobiology and host response; and improvements to canine rabies elimination regionally throughout Africa, Asia, and the Americas by application of the best technical, organizational, economic, and socio-political practices. Significantly, anticipated Gavi support will enable improved access of human rabies vaccines in lesser developed countries at a national level, with integrated bite management, dose-sparing regimens, and a 1 week vaccination schedule.

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“…In our study, both conventional RT-PCR and real-time RT-PCR had lower specificity than in intra-laboratory evaluations. The same observation was made for proficiency testing focusing on sub-Saharan African countries, where 89% sensitivity and 86% specificity were estimated when using conventional RT-PCR [ 43 ]. Several possibilities may explain variations of specificity in RT-PCR techniques like the primer/probe model [ 57 ], the RT-PCR system (one-step vs. two-step) [ 58 ] as well as the enzyme mix or even the thermocycler [ 58 , 59 ].…”

Section: Discussionmentioning

confidence: 98%

“…Another paper has described the results of a Latin America and Caribbean network of 23 laboratories from a FAT proficiency exercise highlighting the need to harmonise this technique with a mean concordance among participants of 81% [ 42 ]. Recently, a study presented the results of 13 sub-Saharan African laboratories taking part in FAT and conventional RT-PCR proficiency tests with respective mean laboratory concordances of 88% and 98% [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

“…The rabies diagnosis proficiency testing exercises presented in the scientific literature have shown that in practice, the accuracy of diagnostic results could be lower than presented in the literature on reference standards [ 41 – 43 ]. The OIE manual indicates that both FAT and PCR techniques provide a reliable rabies diagnosis in 98% to 100% of cases when appropriate conjugate or primers are respectively used, and that FAT is recognised as a highly sensitive and specific technique (between 96% and 99%)[ 44 ].…”

Section: Discussionmentioning

confidence: 99%

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Multi-annual performance evaluation of laboratories in post-mortem diagnosis of animal rabies: Which techniques lead to the most reliable results in practice?

et al. 2021

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Rabies diagnosis proficiency tests on animal specimens using four techniques (FAT, RTCIT, conventional RT-PCR and real-time RT-PCR) were organised over 10 years (2009–2019). Seventy-three laboratories, of which 59% were from Europe, took part. As the panels were prepared with experimentally-infected samples, the error rate of laboratories on positive and negative samples was accurately estimated. Based on fitted values produced by mixed modelling including the variable “laboratory” as a random variable to take into account the longitudinal design of our dataset, the technique that provided the most concordant results was conventional RT-PCR (99.3%; 95% CI 99.0–99.6), closely followed by FAT (99.1%; 95% CI 98.7–99.4), real-time RT-PCR (98.7%; 95% CI 98.1–99.3) and then RTCIT (96.8%; 95% CI 95.8–97.7). We also found that conventional RT-PCR provided a better diagnostic sensitivity level (99.3% ±4.4%) than FAT (98.7% ±1.6%), real-time RT-PCR (97.9% ±0.8%) and RTCIT (95.3% ±5.1%). Regarding diagnostic specificity, RTCIT was the most specific technique (96.4% ±3.9%) followed closely by FAT (95.6% ±3.8%), real-time RT-PCR (95.0% ±1.8%) and conventional RT-PCR (92.9% ±0.5%). Due to multiple testing of the samples with different techniques, the overall diagnostic conclusion was also evaluated, and found to reach an inter-laboratory concordance level of 99.3%. The concordance for diagnostic sensitivity was 99.6% ±2.0% and for diagnostic specificity, 98.0% ±8.5%. Molecular biology techniques were, however, found to be less specific than expected. The potential reasons for such findings are discussed herein. The regular organisation of performance tests has contributed to an increase in the performance of participating laboratories over time, demonstrating the benefits of such testing. Maintaining a high-quality rabies diagnosis capability on a global scale is key to achieving the goal of eliminating dog-mediated human rabies deaths. The regular organisation of exercises on each continent using selected local strains to be tested according to the local epidemiological situation is one factor that could help increase reliable diagnosis worldwide. Rabies diagnosis capabilities could indeed be enhanced by providing adequate and sustainable proficiency testing on a large scale and in the long term

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

Cited by 8 publications

References 33 publications

Evaluation of the diagnostic accuracy of lateral flow devices as a tool to diagnose rabies in post-mortem animals

Evaluation of the diagnostic accuracy of lateral flow devices as a tool to diagnose rabies in post-mortem animals

Update on lyssaviruses and rabies: will past progress play as prologue in the near term towards future elimination?

Multi-annual performance evaluation of laboratories in post-mortem diagnosis of animal rabies: Which techniques lead to the most reliable results in practice?

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