Comparison of a competitive inhibition ELISA and the card agglutination test for detection of antibodies to Anaplasma marginale and Anaplasma centrale in cattle

Molloy, J.B.; Bowles, P.M.; Knowles, Donald P.; McElwain, Terry F.; Bock, R.E.; Kingston, T G; Blight, GW; Dalgliesh, R.J.

doi:10.1111/j.1751-0813.1999.tb11712.x

Cited by 29 publications

(32 citation statements)

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“…10,17,23,26,28,31 To the authors' knowledge, this is the first attempt at estimating accuracy of these tests using a Bayesian latent class approach. The MSP-5 cELISA had the highest accuracy of the 3 evaluated tests based on the dichotomous interpretation with a Youden index of 0.88.…”

Section: Discussionmentioning

confidence: 99%

“…Diagnostic accuracy of the card test, MSP-5 cELISA, and qPCR were estimated in a latent class analysis using a Bayesian framework (evaluation of direct smears were excluded because of few positive tests). Prior probability distributions for this analysis (Table 1) were based on published literature 26,31 and modified by expert opinion because previous studies employed different reference standards. A 3-test, single population model was used assuming conditional independence between pairs of diagnostic tests.…”

Section: Discussionmentioning

confidence: 99%

“…Percent inhibition was calculated as (mean OD of negative control -mean OD of test sera)/(mean OD of negative control) 3 100, and results were interpreted at a cutoff of 30% inhibition. 26 Samples with greater than or equal to 30% inhibition were considered positive.…”

Section: Diagnostic Testingmentioning

confidence: 99%

“…2,35 The diagnostic sensitivity (Se) and specificity (Sp) of the card test has been estimated as 98% and 99%, respectively. 26 The diagnostic accuracy of the MSP-5 cELISA estimated relative to a nested PCR coupled with MSP-5 sequence analysis and hybridization at a cutoff point of 28% inhibition had a relative Se of 96% and Sp of 95%. 31 From the Department of Veterinary Integrative Biosciences, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX (Fosgate, Dobek); and the Departments of Large Animal Clinical Sciences (Urdaz-Rodrí-guez, Rae, Donovan, Melendez) and Physiological Sciences (Dunbar, Alleman), College of Veterinary Medicine, University of Florida, Gainesville, FL.…”

Section: Introductionmentioning

confidence: 99%

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Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

et al. 2010

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Abstract. Bovine anaplasmosis (BA) is a hemoparasitic disease of great importance in cattle within the tropical and subtropical regions of the world. Control programs for BA require accurate diagnostic assays but validation can be challenging because the true disease status of all animals is frequently not known with certainty. The objective of this study was to estimate the accuracy of assays for detection of Anaplasma marginale infection in lactating dairy cattle of Puerto Rico using Bayesian methods without a perfect reference test. There were 2,331 cattle with complete diagnostic results sampled from 79 herds, and the prevalence of BA was estimated as 22% (95% probability interval [PI]: 19-25%). The sensitivity (Se) and specificity (Sp) of a major surface protein 5 competitive enzyme-linked immunosorbent assay (MSP-5 cELISA) were estimated as 99% (95% PI: 96-100%) and 89% (95% PI: 87-92%), respectively. The Se and Sp of a quantitative polymerase chain reaction (qPCR) were 67% (95% PI: 60-74%) and 99% (95% PI: 99-100%). The Se and Sp of a card agglutination test were 34% (95% PI: 29-39%) and 99% (95% PI: 99-100%). Area under the receiveroperating characteristic curve for the MSP-5 cELISA was 0.748 (95% PI: 0.71-0.79). The MSP-5 cELISA appears to be the test of choice for screening cattle for subclinical BA based on the high estimated Se, rapidity of results, relative low cost, and ease of standardization.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Diagnostic Testingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

et al. 2010

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“…The IFA test, which did not discriminate between Anaplasma species because of antigenic similarity (Palmer, 1989 andVisser et al, 1992) were widely used for serodiagnosis of anaplasmosis (Goff et al, 1985, Montenegro-James et al, 1985and Shkap et al, 1990. A commercial cELISA based on the MSP 5 surface protein has been developed for detection of antibodies against A. marginale (Knowles et al, 1996 andTorioni de Echaide et al, 1998) but, similarly to the IFA, it was found not to be applicable for discrimination between A. centrale-specific antibodies (Molloy et al, 1999 andDe la Fuente et al, 2005). The IFA based on 80 kDa A. centrale antigen differentiated between A. marginale and A. centrale in patent infections only, while the cELISA based on 116 kDa antigen was highly sensitive and specific in carrier infections (Molloy et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

Molecular and serological detection of A. centrale- and A. marginale-infected cattle grazing within an endemic area

Molad¹,

Mazuz²,

Fleiderovitz³

et al. 2006

Veterinary Microbiology

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A reverse line blot hybridization (RLB) one-stage nested PCR (nPCR) for Anaplasma centrale and a nested PCR for Anaplasma marginale were used to detect infected cattle grazing within an endemic region in Israel. A novel set of PCR primers and oligonucleotide probes based on a 16S ribosomal RNA gene was designed for RLB detection of both Anaplasma species, and the performance of the molecular assays compared. The immunofluorescent antibody test (IFA) was used to detect antibodies to both Anaplasma species, whereas, a highly sensitive and specific competitive enzymelinked immunosorbent assay (cELISA) was used to detect antibodies in A. centralevaccinated cattle. The RLB and the nested PCR procedures showed bacteremia with sensitivity of 50 infected erythrocytes per milliliter. Up to 93% of the A. centrale vaccinates carried specific antibodies that were detected by cELISA, and up to 71% of the vaccinated cattle were found to be naturally infected with A. marginale according to the PCR and the RLB assays. Nevertheless, no severe outbreaks of A. marginale infection occurred among vaccinated herds in this endemic region. It appears that both, molecular tools and serology are useful for evaluation of the vaccine efficacy. In the light of wide natural field infection with A. marginale, strong recommendations to continue the A. centrale vaccination program regime will continue until a new generation of non-bloodbased vaccine will be developed. Article Outline

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Detection of the Anaplasma centrale vaccine strain and specific differentiation from Anaplasma marginale in vaccinated and infected cattle

Shkap¹,

Molad²,

Fish³

et al. 2002

Parasitol Res

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Bovine anaplasmosis caused by the intraerythrocytic rickettsia Anaplasma marginale is the most prevalent tick-borne disease of cattle worldwide. The most efficient method to control anaplasmosis is by vaccination using live Anaplasma centrale, a closely related species or subspecies of low pathogenicity that is capable of inducing significant protection against the more virulent A. marginale. In the present study, we applied PCR assays to detect and discriminate field infection with A. marginale from A. centrale persistently infected vaccinates. Direct and one-stage nested PCR were based on A. centrale mbp58 specific sequence, with the assay sensitivity level of 0.00001% for nested PCR performed in a single amplification step. Size polymorphism in the A. marginale msp1 alpha gene among strains was used to design a PCR capable of discriminating between the Israel T and NT strains of A. marginale and the encoded MSP1a size polymorphism was confirmed by immunoprecipitation. The detection of A. centrale in 72% of vaccinated field-grazing cattle clearly indicated that the majority of vaccinated cattle remain carriers. A. marginalewas detected in 64% of these vaccinated cattle, demonstrating that, as expected, natural transmission occurs within the endemic region. The lack of severe A. marginaleoutbreaks in this region, despite ongoing transmission, is consistent with protection being provided by widespread vaccination with A. centrale.

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Comparison of a competitive inhibition ELISA and the card agglutination test for detection of antibodies to Anaplasma marginale and Anaplasma centrale in cattle

Abstract: We conclude that the competitive inhibition ELISA is a useful alternative to the card agglutination test for detection of A marginale or A centrale infection in cattle. The assay should be particularly useful for epidemiological applications such as prevalence studies and control programs.

Cited by 29 publications

References 5 publications

Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

Diagnostic Accuracy of Methods for Detecting Anaplasma Marginale Infection in Lactating Dairy Cattle of Puerto Rico

Molecular and serological detection of A. centrale- and A. marginale-infected cattle grazing within an endemic area

Detection of the Anaplasma centrale vaccine strain and specific differentiation from Anaplasma marginale in vaccinated and infected cattle

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