Fluorescence polarization assay for the diagnosis of bovine brucellosis: adaptation to field use

Nielsen, Klaus; Gall, D.; Smith, Phillip D.; Kelly, William A.; Yeo, Jennifer; Kenny, Kevin; Heneghan, Terry; McNamara, Sharon; Maher, Peter; O'Connor, Jane M.; Walsh, Bradley J.; Carroll, J.; Rojas, X; Rojas, Fresia; Pérez, Bárbaro Agustín Armas; Wulff, O.; Buffoni, L.; Salustio, E.; Gregoret, R.; Samartino, Luis; Dajer, A; Luna-Martinez, E.

doi:10.1016/s0378-1135(00)00386-2

Cited by 30 publications

(24 citation statements)

References 8 publications

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“…As provas sorológicas empregadas no diagnóstico são teste de triagem do Antígeno Acidificado Tamponado (AAT), Fixação do Complemento (FC) e teste de Polarização Fluorescente (PF) (Nielsen 2001). Porém, podem ser empregados outros testes como o imunoenzimático competitivo (ELISA-C) (Paulin et al 2012) e ensaio imunoenzimático com proteínas-G (ELISA-G) (Kumar & Chand 2011).…”

Section: Infecção Transplacentária E Intrauterina Porunclassified

Infecção transplacentária e intrauterina por Brucella abortus em búfalos (Bubalus bubalis)

et al. 2015

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RESUMO: O objetivo deste trabalho foi verificar a presença de Brucella abortus e as lesões causadas por esse agente nos anexos fetais e nos fetos de búfalas. Para isso, 20 búfalas em diversos meses de gestação, sorologicamente positivas para brucelose, foram submetidas ao abate sanitário. A idade fetal foi determinada através de exames ultrassonográficos associados à mensuração dos fetos durante a necropsia. Do útero fechado desses animais foram coletadas amostras para histopatologia e qPCR. A partir do segundo mês de gestação foi possível detectar a presença de DNA de B. abortus em líquido amniótico, líquido alantoide e em útero e, a partir do quinto mês, na placenta, coração, baço, rim, pulmão, intestino, fígado e linfonodos dos fetos. Os principais achados anatomopatológicos foram placentite fibrinopurulenta necrótica e endometrite supurativa crônica.

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Section: Infecção Transplacentária E Intrauterina Porunclassified

Infecção transplacentária e intrauterina por Brucella abortus em búfalos (Bubalus bubalis)

et al. 2015

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“…The method has been validated for a large number of species, including humans, cattle, and bisons, and is a prescribed OIE test applied for the examination of cattle and pigs during international turnover (19). The various studies concerning bovine sera showed the high sensitivity and specificity of the method (20)(21)(22). Unfortunately, there is a lack of information about the practical aspects and validation of FPA in diagnosis of porcine brucellosis (18,19).…”

Section: Discussionmentioning

confidence: 99%

“…If a molecule is labelled, the rotation can be determined by measuring polarised light intensity in vertical and horizontal planes (19). It is host species-independent method and can be performed also with whole blood or milk (9,11,(20)(21)(22). According to OIE Terrestrial Manual 2009, the FPA is recommended as a prescribed test for bovine and porcine brucellosis for determining the health status of animals (19).…”

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confidence: 99%

Value of Fluorescence Polarisation Assay in Comparison to Traditional Techniques in Diagnosis of Porcine Brucellosis

Weiner

Iwaniak

Szulowski

2012

Bulletin of the Veterinary Institute in Pulawy

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The aim of this study was the evaluation of fluorescence polarisation assay (FPA) in the diagnosis of porcine brucellosis in comparison with Rose Bengal test (RBT), serum agglutination test (SAT), complement fixation test (CFT), 2-mercaptoethanol test, and ELISA. Eight hundred seventeen sera from pigs, including 612 sera from healthy animals, seven sera from Brucella suis bv2 culture positive animals, and 198 sera classified as false positive, originated from confirmatory investigations, were used. All sera from healthy animals, negative in RBT, SAT, CFT, and ELISA were also negative in FPA. All sera positive in serological examination, originated from Brucella infected animals, were also positive in FPA. Among sera classified as false positive almost half of the samples tested (49.49%) reacted positively in FPA. The examinations confirmed the usefulness of FPA in diagnosis of porcine brucellosis, but the method, like the other tests, does not allow resolving the problem of discrimination cross-reacting from specific antibodies.

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“…The technique is highly suitable to high-throughput testing but also presents a valid option for low-throughput tests. The fluorescent mechanism of the technique suggests that it may also be possible to develop tests that use whole-blood samples (29) and milk samples (30), as has been the case for the FPA. Likewise, a portable single-tube assay could be developed.…”

Section: Discussionmentioning

confidence: 99%

Time-Resolved Fluorescent Resonance Energy Transfer Assay for Simple and Rapid Detection of Anti-BrucellaAntibodies in Ruminant Serum Samples

et al. 2009

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Brucellosis is a globally significant zoonosis, the control of which is difficult and resource intensive. Serological tests form a vital part of a multifactorial approach to control and are often performed in large numbers. The aim of the present study was to develop a new assay to improve the efficiency, ease, and effectiveness of serological testing. An existing competitive enzyme-linked immunosorbent assay (cELISA) was adapted to a completely homogeneous time-resolved fluorescent resonance energy transfer (TR-FRET) assay. This was achieved by labeling an anti-Brucella monoclonal antibody with a long-lifetime donor fluorophore and Brucella smooth lipopolysaccharide with a compatible acceptor and optimizing the reading conditions. The assay was performed in a 96-well plate with a single 30-min incubation period and no separation (wash) steps and was concluded by a single plate-reading step. The performance of the assay was evaluated with a panel of serum samples from infected (n ‫؍‬ 73) and uninfected (n ‫؍‬ 480) sources and compared to the performance of the parent cELISA, an indirect ELISA (iELISA), and fluorescence polarization assay (FPA). The performance of the TR-FRET assay matched the performance of the iELISA, which had 100% diagnostic sensitivity and specificity, and surpassed the performance of the cELISA and the FPA. The results also demonstrated that the TR-FRET technique is effective with poor-quality serum samples from the field. To the knowledge of the authors, this is the first homogeneous TR-FRET assay to detect antibodies raised against an infectious disease. The technique appears to be sufficiently adaptable to meet the needs of many other similar testing requirements to identify infectious diseases.Brucellosis is a zoonosis of widespread distribution and significance caused by species of the genus Brucella. The disease is known to be especially prevalent in the Middle East and Mediterranean Basin (33), and there are disturbing signs of its reemergence across large areas of the globe, especially in central Asia. Few countries have successfully managed to eradicate the disease. In many of these countries, although the livestock sector is disease free, a significant wildlife reservoir remains and presents a risk of reintroduction (11).The principle etiologic agents of brucellosis are the classical smooth species Brucella abortus, B. melitensis, and B. suis. These species all have smooth lipopolysaccharide (sLPS), which is a major virulence factor (17, 35). Each of these species has a preferred host; however, many animals, including humans, are susceptible to each of the classical species, although the underlying mechanisms for host preference are not clearly understood (5). Brucellosis in ruminants is mainly manifested by reproductive failure due to abortion. There are few other clinical signs, and this causes difficulties with diagnosis. In humans, the disease is mainly presented as an undulating febrile condition, although there may be other, more serious complications. The symptoms of human ...

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Fluorescence polarization assay for the diagnosis of bovine brucellosis: adaptation to field use

Cited by 30 publications

References 8 publications

Infecção transplacentária e intrauterina por Brucella abortus em búfalos (Bubalus bubalis)

Infecção transplacentária e intrauterina por Brucella abortus em búfalos (Bubalus bubalis)

Value of Fluorescence Polarisation Assay in Comparison to Traditional Techniques in Diagnosis of Porcine Brucellosis

Time-Resolved Fluorescent Resonance Energy Transfer Assay for Simple and Rapid Detection of Anti-BrucellaAntibodies in Ruminant Serum Samples

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