A diagnostic evaluation of real-time PCR, fluorescent antibody and microscopic agglutination tests in cases of equine leptospiral abortion.
Abstract: A comprehensive evaluation of the real-time PCR assay for leptospirosis in comparison with other diagnostic assays on a large-scale basis is fundamental in validating the assay and determining the causes of equine abortions. Objective: To compare and evaluate the diagnostic value of real-time PCR assay for leptospirosis with traditional methods in equine leptospiral abortions. Methods: Cross-sectional observational study. Methods: A Leptospira spp. fluorescent antibody test (FAT), microscopic agglutination test (MAT) and real-time PCR (targeting the LipL32 gene) were compared and evaluated in equine fetal necropsy specimens (placenta, kidney, liver and heart blood) and maternal serum (when available) in 339 equine fetuses. Results: From a total of 339 equine fetuses necropsied, 21 cases (6.19%) were diagnosed as leptospiral abortion. The majority of leptospiral abortions occurred in January (8 cases) and February (5 cases). Real-time PCR detected 21 of 21 cases, whereas MAT and FAT detected 19 and 18 (including 2 suspicious cases) cases, respectively. Comparing tissues, placenta yielded somewhat similar cycle of threshold values by real-time PCR compared with kidney, whereas kidney was the best specimen for the diagnosis of leptospirosis by the FAT test. In all MAT positive cases, the predominant titre in fetal heart blood was to serovar Pomona (ranging 1:100 to 1:204,800) with little or no cross-reaction to serovar Grippotyphosa. Conclusions: The results indicate that real-time PCR is an effective method for the diagnosis of leptospiral abortion in horses. However, MAT should continue to be used in clinical cases for serovar determination.
© 2014 EVJ Ltd.
Publication Date: 2014-05-23 PubMed ID: 24750279DOI: 10.1111/evj.12281Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study explores the use of real-time PCR assay for diagnosing cases of equine leptospiral abortion, comparing it with traditional methods like the fluorescent antibody test (FAT) and microscopic agglutination test (MAT). The research suggests that real-time PCR is more effective for diagnosing leptospiral abortions in horses.
Research Methods
- The study is a cross-sectional observational one that uses multiple diagnostic testing methods, including real-time PCR, FAT, and MAT.
- The researchers tested placenta, kidney, liver, and heart blood samples from 339 equine fetuses. They also used maternal serum where available.
- The real-time PCR targeted a specific gene known as LipL32.
Results
- Out of the 339 equine fetuses examined, 21 cases (representing 6.19%) were diagnosed as leptospiral abortion.
- These abortions mostly happened in the months of January (8 cases) and February (5 cases).
- Real-time PCR was able to detect all 21 cases, while MAT and FAT detected 19 and 18 cases, respectively.
- When comparing tissue samples, the placenta yielded comparable cycle threshold values to the kidney with real-time PCR, but the kidney proved to be the best specimen for the FAT test.
- In all MAT-positive cases, the predominant titre in fetal heart blood was to serovar Pomona, with little or no cross-reaction to serovar Grippotyphosa.
Conclusions
- Based on the findings of this study, real-time PCR is a highly effective method for diagnosing leptospiral abortion in horses.
- However, the researchers recommend that the MAT continues to be used in clinical cases for the determination of different types of serovars.
Cite This Article
APA
Erol E, Jackson CB, Steinman M, Meares K, Donahoe J, Kelly N, Locke S, Smith JL, Carter CN.
(2014).
A diagnostic evaluation of real-time PCR, fluorescent antibody and microscopic agglutination tests in cases of equine leptospiral abortion.
Equine Vet J, 47(2), 171-174.
https://doi.org/10.1111/evj.12281 Publication
Researcher Affiliations
- Veterinary Diagnostic Laboratory, University of Kentucky, Lexington, USA.
MeSH Terms
- Abortion, Veterinary / diagnosis
- Abortion, Veterinary / microbiology
- Agglutination Tests / methods
- Agglutination Tests / veterinary
- Animals
- Cross-Sectional Studies
- Female
- Fluorescent Antibody Technique
- Horse Diseases / diagnosis
- Horse Diseases / microbiology
- Horse Diseases / pathology
- Horses
- Leptospira / isolation & purification
- Leptospirosis / complications
- Leptospirosis / veterinary
- Pregnancy
- Pregnancy Complications, Infectious / veterinary
- Real-Time Polymerase Chain Reaction / veterinary
Citations
This article has been cited 7 times.- Lei C, Liao K, Zhao Y, Long Z, Zhu S, Wu J, Xiao M, Zhou J, Zhang S, Li L, Zhu Y, Lu D, Yang J, Sun X. A Novel System for the Detection of Spontaneous Abortion-Causing Aneuploidy and Its Erroneous Chromosome Origins through the Combination of Low-Pass Copy Number Variation Sequencing and NGS-Based STR Tests. J Clin Med 2023 Feb 23;12(5).
- Aras Z, Yavuz O. Evaluation of fetal heart serum amyloid a concentrations in infectious cattle abortion cases. Heliyon 2022 Nov;8(11):e11330.
- Fagre AC, Mayo CE, Pabilonia KL, Landolt GA. Seroprevalence of Leptospira spp. in Colorado equids and association with clinical disease. J Vet Diagn Invest 2020 Sep;32(5):718-721.
- Fouché N, Graubner C, Lanz S, Schweighauser A, Francey T, Gerber V. Acute kidney injury due to Leptospira interrogans in 4 foals and use of renal replacement therapy with intermittent hemodiafiltration in 1 foal. J Vet Intern Med 2020 Mar;34(2):1007-1012.
- Trimble AC, Blevins CA, Beard LA, Deforno AR, Davis EG. Seroprevalence, frequency of leptospiuria, and associated risk factors in horses in Kansas, Missouri, and Nebraska from 2016-2017. PLoS One 2018;13(10):e0206639.
- Filipe J, Lauzi S, Bullo F, D'Incau M, Meroni G, Martino PA, Magistrelli S, Restelli M, Dall'Ara P. Leptospira spp. Antibody Seroprevalence in Stray Dogs and Cats: A Study in Milan, Northern Italy. Vet Sci 2024 Oct 5;11(10).
- Zečević I, Picardeau M, Vince S, Hađina S, Perharić M, Štritof Z, Stevanović V, Benvin I, Turk N, Lohman Janković I, Habuš J. Association between Exposure to Leptospira spp. and Abortion in Mares in Croatia. Microorganisms 2024 May 21;12(6).
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