Improving the laboratory diagnosis of leptospiral uveitis in horses by using an indirect ELISA for the detection of antibodies against Leptospira spp. in intraocular samples

Loibl, J.; Gerhards, H; Brem, S; Wollanke, B

doi:10.21836/pem20180308

Cited by 11 publications

(25 citation statements)

References 35 publications

(53 reference statements)

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“…It could be shown in earlier investigations that ELISA testing was more sensitive compared to MAT in detecting intraocular anti-leptospiral antibodies (Loibl 2009, Loibl et al 2018. A laboratory ELISA quick test (SNAP ® Lepto, Fa.…”

Section: Introductionmentioning

confidence: 99%

Evaluation of “SNAP® Lepto”-ELISA and comparison with MAT and PCR results for diagnosis of leptospiral uveitis in horses using intraocular samples

Wollanke¹,

Geiger²,

Gerhards³

2018

PHK

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Chronic intraocular leptospiral infection is the most common cause for recurrent uveitis in equids. History and a combination of typical ophthalmological findings can lead to suspected diagnosis of leptospiral induced uveitis. Testing intraocular fluids had been shown to be the most valuable tool for providing evidence of an intraocular leptospiral infection. The most appropriate treatment for recurrences of leptospiral uveitis is vitrectomy, whereas vitrectomy in non-leptospiral uveitis cases is less successful in regard of avoiding further damage to effected eyes. However, in cases in which clinical and ophthalmological findings are not typical for leptospiral induced uveitis, preoperative testing of aqueous fluid can be indicated to evaluate an indication for vitrectomy. Until now, for save sampling of aqueous humour, a general anaesthesia was necessary for paracentesis of the anterior chamber, and a second was necessary for vitrectomy in case of leptospira-positive laboratory results (either antibodies using microscopic agglutination tests (MAT), enzyme-linked immunosorbent assays (ELISA) or polymerase chain reactions (PCR) detecting LipL32). A quick ELISA test, the SNAP ® Lepto, has recently been made available to detect antibodies directed against LipL32 of pathogenic leptospires in any species. In order to evaluate the SNAP ® Lepto and to test whether it is effective with intraocular samples (aqueous and vitreous humour) from horses, 118 samples from equine eyes suffering from recurrent uveitis and 120 samples from equine control eyes were taken and tested with SNAP ® Lepto (detecting antibodies directed against LipL32), MAT (detecting antibodies directed against ten different serovars) and PCR (detecting the LipL32 antigen). If one of these tests was positive, an intraocular leptospiral infection was assumed.The result was that SNAP ® Lepto was positive in 114 out of 118 intraocular samples (97 %) from horses suffering from recurrent uveitis. SNAP ® Lepto was even superior to MAT, with a sensitivity of 0.97 (MAT 0.93). PCR was positive with fluids from eyes suffering from uveitis in 70 % of samples. There were no positive PCR results in any SNAP ® Lepto-negative samples from uveitic eyes and the MAT was positive in four SNAP ® Lepto-negative samples. On the other hand, SNAP ® Lepto gave a positive result in eight MAT-negative samples from eyes with uveitis. Furthermore, none of the antibody tests was positive regarding any sample from the control eyes, and PCR was positive in one single sample from a control eye. It can be concluded that SNAP ® Lepto, which gives a result within 10 minutes, is a valuable test method for detecting antibodies directed against LipL32 in intraocular fluids from horses. In the case of a negative result, MAT and PCR can be performed subsequently to reduce false negative results. Keywords: chronic intraocular leptospiral infection, equine recurrent uveitis (ERU), MAT, ELISA, serum, SNAP ® Lepto, anterior chamber fluid, vitreous Citation: Wollanke B., Geiger T., Gerhards H. (2018...

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Section: Introductionmentioning

confidence: 99%

Evaluation of “SNAP® Lepto”-ELISA and comparison with MAT and PCR results for diagnosis of leptospiral uveitis in horses using intraocular samples

Wollanke¹,

Geiger²,

Gerhards³

2018

PHK

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“…For example, anti-Leptospira antibodies are regularly detected in vitreous material obtained during vitrectomies [13,15,21,[32][33][34][35][36][37][38][39][40][41]. These antibodies are detected using the micro agglutination test (MAT), and various enzyme-linked immunosorbent assays (ELISA) [42,43]. In other studies, Leptospira spp.…”

Section: Introductionmentioning

confidence: 99%

“…In addition to direct evidence of the leptospiral infection of vitreous samples from eyes affected with ERU, evidence of autoimmunity also exists [49]. However, Prof. Deeg's research group [50] performed all investigations using vitreous samples, from which leptospiral infections were also regularly detected [13,21,39,[41][42][43]46]. Furthermore, while vitrectomy removes the vitreous material, potential autoantigens from other tissues -especially the lens and retina-remain in the eye [18,39] which contradicts the idea of autoimmunity [39,51].…”

Section: Introductionmentioning

confidence: 99%

In Vivo Biofilm Formation of Pathogenic Leptospira spp. in the Vitreous Humor of Horses with Recurrent Uveitis

et al. 2021

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Equine recurrent uveitis (ERU) causes painful inflammatory attacks and oftentimes blindness in the affected eyes. The disease is considered a late sequela of systemic leptospirosis. The most effective therapy is the surgical removal of the vitreous (vitrectomy), which is not only therapeutic, but provides vitreous material that can be assessed diagnostically. For example, the lipL32 gene, culturable Leptospira spp., and anti-Leptospira antibodies have all been detected in vitreous samples obtained from eyes with chronic ERU. Despite this clear evidence of leptospiral involvement, the systemic administration of antibiotics in infected horses is ineffective at resolving ERU. This syndrome of chronic recurrent inflammation, which is unresponsive to antibiotic therapy, combined with apparent bacteria evading the immune response, is consistent with a biofilm-associated infection. The purpose of this study, therefore, was to detect the in vivo biofilm formation of Leptospira spp. in vitreous samples collected during vitrectomy and examined using a Warthin-Starry silver stain and immunohistochemistry. All known steps of biofilm formation were visualized in these samples, including individual Leptospira spp., leptospiral microcolonies and dense roundish accumulations of Leptospira spp. In many instances spirochetes were surrounded by an extracellular substance. Taken together, data from the present study show that ERU is a biofilm-associated intraocular leptospiral infection, which best explains the typical clinical course.

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“…Therefore, an antibody titer in a serum sample determined with MAT has no significance for the diagnosis of ERU in an individual horse [1,[3][4][5][6]10,[62][63][64][65]. Consequently, serum tests using MAT do not allow a careful decision on the indication for surgery.…”

Section: Introductionmentioning

confidence: 99%

“…For this reason, aqueous humor testing is indicated preoperatively in questionable ERU cases [5,6,11,[65][66][67][68][69]. If either anti-Leptospira antibodies are detectable in the aqueous humor and/or the LipL32 gene of pathogenic Leptospira spp.…”

Section: Introductionmentioning

confidence: 99%

Detection of Anti-LipL32 Antibodies in Serum Samples from Horses with Chronic Intraocular Infection with Leptospira spp.

2021

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Equine recurrent uveitis (ERU) is typically caused by chronic intraocular leptospiral infection in warm-blooded horses in central Europe. The most effective therapy for leptospiral-induced ERU is the surgical removal of diseased vitreous (vitrectomy). Since vitrectomy is a highly specialized and invasive surgery, the indication must be determined very carefully. In order to obtain evidence of intraocular leptospiral infection by laboratory diagnostics in questionable leptospiral ERU-cases, sampling of aqueous humor is required, because serum tests using microscopic agglutination test (MAT) are too unspecific. The SNAP Lepto is a cross-species rapid test for the detection of anti-Lipl32 antibodies that has a high sensitivity (0.97) and specificity (1.00) for the detection of anti-leptospiral antibodies using aqueous humor or vitreous samples, which is comparable to MAT. To evaluate sensitivity and specificity of SNAP Lepto using serum, serum samples from 90 horses with confirmed leptospiral ERU and from 103 ocularly healthy horses were tested by both MAT and SNAP Lepto. Sensitivity was similar for both tests (0.82 vs. 0.79), but specificity was lower for MAT (0.52 vs. 0.95). Sensitivity and specificity are therefore lower in serum samples compared to intraocular samples, however, the SNAP Lepto is far superior to MAT and suitable as a screening method using equine serum.

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Improving the laboratory diagnosis of leptospiral uveitis in horses by using an indirect ELISA for the detection of antibodies against Leptospira spp. in intraocular samples

Cited by 11 publications

References 35 publications

Evaluation of “SNAP® Lepto”-ELISA and comparison with MAT and PCR results for diagnosis of leptospiral uveitis in horses using intraocular samples

Evaluation of “SNAP® Lepto”-ELISA and comparison with MAT and PCR results for diagnosis of leptospiral uveitis in horses using intraocular samples

In Vivo Biofilm Formation of Pathogenic Leptospira spp. in the Vitreous Humor of Horses with Recurrent Uveitis

Detection of Anti-LipL32 Antibodies in Serum Samples from Horses with Chronic Intraocular Infection with Leptospira spp.

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