Anaplasma marginale: Detection of carrier cattle by PCR-ELISA

Gale, K. R.; Dimmock, Christine M.; Gartside, Michael G.; Leatch, G.

doi:10.1016/s0020-7519(96)80009-9

Cited by 35 publications

(20 citation statements)

References 16 publications

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“…However, the rickettsemia of chronically infected animals is generally low, hindering its diagnosis (Palmer et al 1986). In this context, molecular diagnostic assays such as PCR have been developed and are both highly sensitive and specific (Palmer et al 1986, Gale et al 1996, Corona et al 2000.…”

Section: Introductionmentioning

confidence: 99%

Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Bacanelli

Ramos

Araújo³

2014

Pesq. Vet. Bras.

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Pesq. Vet. Bras. 34(1):29-33, janeiro 2014 29 RESUMO.-[Diagnóstico molecular de Anaplasma marginale em bovinos: avaliação quantitativa de uma PCR em tempo real baseada no gene msp5.] A riquétsia Anaplasma marginale é considerada o principal agente da anaplasmose bovina. Devido a não especificidade dos sinais clínicos, a confirmação da infecção nos animais depende de testes laboratoriais. Recentemente, métodos de diagnóstico molecular têm sido aplicados para detecção de A. marginale em bovinos. No entanto, a grande quantidade de testes com diferentes sensibilidade e custos tem dificultado a escolha do ensaio mais adequado. No presente estudo, uma PCR em tempo real baseada no gene msp5 foi avaliada quantitativamente e comparada a uma reação de PCR convencional. As reações foram submetidas à avaliação de sensibilidade e especificidade com DNA plasmidial e amostras provenientes de bovinos experimentalmente infectados por A. marginale. Uma avaliação comparativa a campo foi realizada entre os testes utilizando amostras provenientes de bovinos criados em uma região de estabilidade enzoótica para A. marginale. Embora os testes não tenham apresentado diferença estatisticamente significativa, a PCR em tempo real apresentou valor de sensibilidade maior do que a PCR convencional. A PCR em tempo real foi capaz de detectar uma cópia de msp5 em 100ng de DNA plasmidial, e mais de 80% de resultados positivos entre bovinos experimentalmente infectados apenas sete dias após infecção. Além disso, baseado em análise in silico, a PCR em tempo real avaliada aqui pode ser útil para detecção de Anaplasma ovis. The rickettsia Anaplasma marginale is considered the main agent of bovine anaplasmosis. Due the nonspecific clinical signs of the anaplasmosis, the diagnosis of infection depends of laboratory confirmation. In recent years, molecular diagnostic methods have been used to detect A. marginale in cattle. However, the existence of a large number of assays of different sensitivity and cost makes the choice of an appropriate test difficult. In the present study, a real-time Polymerase Chain Reaction (PCR) based on the msp5 target gene was quantitatively assessed and compared to an end point PCR. Both reactions were subjected to sensitivity and specificity evaluation using plasmid DNA and samples from cattle experimentally infected with A. marginale. A comparative field trial of the tests was carried out using samples of cattle from a stable enzootic area for A. marginale. The real-time PCR showed a higher sensitivity than the end point PCR. This reaction (i.e. real-time PCR) was able to detect one copy of the msp5 gene in 100 ηg of plasmidial DNA, and more than 80% of its results were positive among experimentally infected animals seven days after infection. In addition, based on in silico analysis, the real-time PCR evaluated in the present study appears to be useful for the detection of A. ovis.

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Section: Introductionmentioning

confidence: 99%

Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Bacanelli

Ramos

Araújo³

2014

Pesq. Vet. Bras.

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“…Diagnosis of the disease is usually based on microscopic examination of stained blood films; however this method can only detect levels of 10 6 infected erythrocytes per ml (Gale et al, 1996) in acute infection but it is not sufficiently sensitive or specific to detect chronic carriers. A cELISA was developed based on antibody binding to recombinant A. marginale major surface protein 5 (msp5) (Knowles et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Molecular and serological prevalence of Anaplasma marginale in cattle of North Central Morocco

Hamou

Rahali

Sahibi

et al. 2012

Research in Veterinary Science

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“…However this conventional method have some disadvantages and can only detect levels of 10 6 infected erythrosites per ml in acute infectons [19] . A. marginale and A. centrale multiplies within red blood cell of infected host, resulting in extravascular hemolysis and anemia.…”

Section: Discussionmentioning

confidence: 99%

Klinik Olarak Anaplasmosis Şüphesi Olan Ruminantlarda cELISA’nın Tanısal Değerinin Belirlenmesi

Selçuk¹,

Alver²,

Çatik³

et al. 2015

Kafkas Univ Vet Fak Derg

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The aim of this study was to determine diagnostic value of cELISA in anaplasmosis in clinically suspected animals and to compare the cELISA results with the clinical examination results. For this purpose a total of 720 ruminants (457 cattle, 146 sheep, 117 goat) were examined in terms of clinical signs. Eighty-eight ruminants consisting of 61 cattle, 11 sheep and 16 goat which had the symptoms of anemia, fever, icterus, weakness, depression and lack of appetite were selected for the study. Blood samples were collected from the jugular vein of all clinically suspected animals and serum samples were separated. A commercially available competitive enzyme-linked immunosorbent assay (C-ELISA) kit was used for determine antibodies to Anaplasma species. cELISA based diagnosis revealed that 47 of 88 serum samples (53.4%) were positive for anaplasmosis. In serological examination Anaplasma specific antibodies were determined in 45.9% of cattle, 63.6% of sheep and 56.2% of goats. Seropositivity rate was statistically differ among the age groups of cattle and the highest seropositvity rate was found in <12 month age (P<0.005). However no difference was found in the seropositivity rate of Anaplasma in sheep and goat in relation to age group. From the data obtained in this study it can be concluded that clinical findings are not sufficient criteria for the diagnosis of anaplasmosis and must be supported by serological examination.

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Anaplasma marginale: Detection of carrier cattle by PCR-ELISA

Cited by 35 publications

References 16 publications

Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Molecular diagnosis of Anaplasma marginale in cattle: quantitative evaluation of a real-time PCR (Polymerase Chain Reaction) based on msp5 gene

Molecular and serological prevalence of Anaplasma marginale in cattle of North Central Morocco

Klinik Olarak Anaplasmosis Şüphesi Olan Ruminantlarda cELISA’nın Tanısal Değerinin Belirlenmesi

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