Diagnosis of foot-and mouth disease by real time reverse transcription polymerase chain reaction under field conditions in Brazil

Paixão, Tatiane Alves da; Neta, Alcina V. Carvalho; Paiva, Naimes O; Reis, Jorge R; Barbosa, Meirivan S; Serra, C; Silva, René R; Beckham, Tammy R.; Martin, Bruce L.; Clarke, Neville P.; Adams, L. Garry; Santos, Renato L.

doi:10.1186/1746-6148-4-53

Cited by 32 publications

(38 citation statements)

References 21 publications

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“…However, Ag‐LFDs have only been validated for use with epithelial suspension and vesicular fluid, and limited analytical sensitivity restricts their usefulness to the acute stage of infection (Ferris et al., , ). As a consequence, attempts to transfer highly sensitive rRT‐PCR assays onto portable detection platforms have been investigated (Hearps et al., ; Howson et al., ; Madi et al., ; Monwina et al., ; Paixão et al., ), but are either not commercially available or are only suitable for research purposes (i.e., not diagnostic use). This study evaluates the performance of a commercially available, lyophilized FMDV pan‐serotype‐specific assay, in combination with a commercially available portable thermocycler, in laboratory and East African field settings.…”

Section: Discussionmentioning

confidence: 99%

“…The compatibility of an OIE-recommended rRT-PCR assay with POCT platforms has already been demonstrated (Hearps, Zhang, & Alexandersen, 2002;Madi et al, 2012;Monwina, Clavijo, Li, Collignon, & Kitching, 2007;Paixão et al, 2008). For instance, portable rRT-PCR has been deployed in field settings for accurate detection of FMDV in serum, epithelial suspensions and oesophageal-pharyngeal fluid (OP) fluid (Howson et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

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Direct detection and characterization of foot-and-mouth disease virus in East Africa using a field-ready real-time PCR platform

Howson

Armson

Lyons

et al. 2017

Transbound Emerg Dis

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SummaryEffective control and monitoring of foot‐and‐mouth disease (FMD) relies upon rapid and accurate disease confirmation. Currently, clinical samples are usually tested in reference laboratories using standardized assays recommended by The World Organisation for Animal Health (OIE). However, the requirements for prompt and serotype‐specific diagnosis during FMD outbreaks, and the need to establish robust laboratory testing capacity in FMD‐endemic countries have motivated the development of simple diagnostic platforms to support local decision‐making. Using a portable thermocycler, the T‐COR™ 8, this study describes the laboratory and field evaluation of a commercially available, lyophilized pan‐serotype‐specific real‐time RT‐PCR (rRT‐PCR) assay and a newly available FMD virus (FMDV) typing assay (East Africa‐specific for serotypes: O, A, Southern African Territories [SAT] 1 and 2). Analytical sensitivity, diagnostic sensitivity and specificity of the pan‐serotype‐specific lyophilized assay were comparable to that of an OIE‐recommended laboratory‐based rRT‐PCR (determined using a panel of 57 FMDV‐positive samples and six non‐FMDV vesicular disease samples for differential diagnosis). The FMDV‐typing assay was able to correctly identify the serotype of 33/36 FMDV‐positive samples (no cross‐reactivity between serotypes was evident). Furthermore, the assays were able to accurately detect and type FMDV RNA in multiple sample types, including epithelial tissue suspensions, serum, oesophageal–pharyngeal (OP) fluid and oral swabs, both with and without the use of nucleic acid extraction. When deployed in laboratory and field settings in Tanzania, Kenya and Ethiopia, both assays reliably detected and serotyped FMDV RNA in samples (n = 144) collected from pre‐clinical, clinical and clinically recovered cattle. These data support the use of field‐ready rRT‐PCR platforms in endemic settings for simple, highly sensitive and rapid detection and/or characterization of FMDV.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Direct detection and characterization of foot-and-mouth disease virus in East Africa using a field-ready real-time PCR platform

Howson

Armson

Lyons

et al. 2017

Transbound Emerg Dis

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“…Epithelial tissues, vesicular fluid or oro‐pharyngeal fluid are routinely used for diagnosis of FMD and virus typing (OIE, ). However, oral swab samples from clinically suspect cases and apparently healthy animals have also been used for isolation of FMDV and virus typing using conventional RT‐PCR (Callens et al., ) or FMD diagnosis and virus typing using real‐time RT‐PCR (Paixao et al., ; Jamal et al., ; Jamal and Belsham, ). All the 35 original samples tested negative in ELISA but 31 of them tested positive in pan‐FMDV diagnostic 3D rRT‐PCR assay.…”

Section: Discussionmentioning

confidence: 99%

Genetic Characterization of Serotypes A and Asia-1 Foot-and-mouth Disease Viruses in Balochistan, Pakistan, in 2011

Ullah

Jamal

Romey

et al. 2016

Transbound Emerg Dis

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This study reports characterization of foot-and-mouth disease virus (FMDV) in samples collected from Balochistan, Pakistan. FMDV was detected by pan-FMDV real-time RT-PCR in 31 samples (epithelial and oral swabs) collected in 2011 from clinical suspect cases. Of these, 29 samples were serotyped by serotype-specific real-time RT-PCR assays and were confirmed by sequencing the VP1 coding region. Sixteen samples were found positive for serotype A and eight for serotype Asia-1, whereas five samples were found positive for both serotypes A and Asia-1. Two serotype A positive samples were found positive for two different strains of serotype A FMDV each. Phylogenetic analyses of serotype A FMDVs showed circulation of at least three different sublineages within the A-Iran05 lineage. These included two earlier reported sublineages, A-Iran05 and A-Iran05 , and a new sublineage, designated here as A-Iran05 . This shows that viruses belonging to the A-Iran05 lineage are continuously evolving in the region. Viruses belonging to the A-Iran05 sublineage showed close identity with the viruses circulating in 2009 in Pakistan and Afghanistan. However, viruses belonging to the A-Iran05 detected in Balochistan, Pakistan, showed close identity with the viruses circulating in Kyrgyzstan, Iran and Kazakhstan in 2011 and 2012, showing that viruses responsible for outbreak in these countries have a common origin. Serotype Asia-1 FMDVs reported in this study all belonged to the earlier reported Group-VII (Sindh-08), which is currently a dominant strain in the West Eurasian region. Detection of two different serotypes of FMDV or/and two different strains of the same serotype in one animal/sample shows complexity in occurrence of FMD in the region.

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“…However, success of these tests entirely depends on sample quality determined by timing and handling of samples [1], and a number of outbreaks in the region were not serotyped and/or characterised. All but one of the 13 NRLs relied on antibody ELISAs, most likely because these tests are cheap and suitable for working on many samples, require lower level of biocontainment [22], and neither depend on cell cultures nor on highly sensitive, expensive and service-requiring PCR-equipment [31]. …”

Section: Discussionmentioning

confidence: 99%

“…The phasing out of the CFT in the region demonstrates a move towards more modern methods, which is also evident from the five NRLs already using or introducing PCR. Only three NRLs used VI despite this being about as sensitive as PCR [31,46], most likely for the same reasons as for not using VNT. Equally few NRLs (3) used real time and/or conventional RT-PCR in routine diagnosis, which accords with findings in another endemic country, Brazil, where limited use was attributed to lack of infrastructure, high cost and anticipated problems of maintaining technically complicated and service-demanding PCR machines [31].…”

Section: Discussionmentioning

confidence: 99%

Laboratory capacity for diagnosis of foot-and-mouth disease in Eastern Africa: implications for the progressive control pathway

et al. 2013

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BackgroundAccurate diagnosis is pertinent to any disease control programme. If Eastern Africa is to work towards control of foot-and-mouth disease (FMD) using the Progressive Control Pathway for FMD (PCP-FMD) as a tool, then the capacity of national reference laboratories (NRLs) mandated to diagnose FMD should match this task. This study assessed the laboratory capacity of 14 NRLs of the Eastern Africa Region Laboratory Network member countries using a semi-structured questionnaire and retrospective data from the World Reference Laboratory for FMD annual reports and Genbank® through National Centre for Biotechnology Information for the period 2006–2010.ResultsThe questionnaire response rate was 13/14 (93%). Twelve out of the 13 countries/regions had experienced at least one outbreak in the relevant five year period. Only two countries (Ethiopia and Kenya) had laboratories at biosecurity level 3 and only three (Ethiopia, Kenya and Sudan) had identified FMD virus serotypes for all reported outbreaks. Based on their own country/region assessment, 12/13 of these countries /regions were below stage 3 of the PCP-FMD. Quarantine (77%) and vaccination (54%) were the major FMD control strategies employed. The majority (12/13) of the NRLs used serological techniques to diagnose FMD, seven used antigen ELISA and three of these (25%) also used molecular techniques which were the tests most frequently requested from collaborating laboratories by the majority (69%) of the NRLs. Only 4/13 (31%) participated in proficiency testing for FMD. Four (31%) laboratories had no quality management systems (QMS) in place and where QMS existed it was still deficient, thus, none of the laboratories had achieved accreditation for FMD diagnosis.ConclusionsThis study indicates that FMD diagnostic capacity in Eastern Africa is still inadequate and largely depends on antigen and antibody ELISAs techniques undertaken by the NRLs. Hence, for the region to progress on the PCP-FMD, there is need to: implement regional control measures, improve the serological diagnostic test performance and laboratory capacity of the NRLs (including training of personnel as well as upgrading of equipment and methods, especially strengthening the molecular diagnostic capacity), and to establish a regional reference laboratory to enforce QMS and characterization of FMD virus containing samples.

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Diagnosis of foot-and mouth disease by real time reverse transcription polymerase chain reaction under field conditions in Brazil

Cited by 32 publications

References 21 publications

Direct detection and characterization of foot-and-mouth disease virus in East Africa using a field-ready real-time PCR platform

Direct detection and characterization of foot-and-mouth disease virus in East Africa using a field-ready real-time PCR platform

Genetic Characterization of Serotypes A and Asia-1 Foot-and-mouth Disease Viruses in Balochistan, Pakistan, in 2011

Laboratory capacity for diagnosis of foot-and-mouth disease in Eastern Africa: implications for the progressive control pathway

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