Detection of intraocular leptospiral DNA, antibodies and Leptospira spp. in horses with equine recurrent uveitis in different laboratories

Baake, Eva I.A.; Borstel, M von; Rohn, Karl; Ohnesorge, Bernhard

doi:10.21836/pem20160407

Cited by 9 publications

(30 citation statements)

References 48 publications

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“…In 14 % of the vitreous specimens of horses with suspected leptospiral uveitis by ophthalmological examination leptospiral antibodies could not be detected neither with MAT nor ELI-SA. This observation is consistent with studies of other authors who also failed to detect specific antibodies in all of the intraocular samples from horses suffering from recurrent uveitis (Wollanke et al 1998a,b, Wollanke et al 2001a,b, Wollanke et al 2004a, Brandes et al 2007, Tömördy et al 2010, Wiehen 2012, Kulbrock et al 2013, Baake et al 2016, Dorrego Keiter et al 2016, Dorrego Keiter et al 2017. The different percentages of Leptospira positive and negative samples might be attributable to different methods in case selection and sampling techniques.…”

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confidence: 89%

“…Leptospiral DNA could be detected in 61% of the vitreous samples from horses with ERU, whereas in the control group none showed a positive reaction. Other authors reported on 57 % to 100 % of positive test results in intraocular samples from horses with ERU (Faber et al 2000, Wollanke 2002, Gesell 2004, Wollanke et al 2004a, Brandes et al 2007, Wiehen 2012, Baake et al 2016, Dorrego Keiter et al 2016, Schinagl 2017. The frequent detection of leptospiral DNA in intraocular material of equine eyes with recurrent uveitis confirmed that leptospires play a decisive role in the aetiology and pathogenesis of the disease.…”

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confidence: 98%

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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

Loibl¹,

Gerhards²,

Brem³

et al. 2018

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Equine recurrent uveitis (ERU) is a frequently occurring disease, at least in horses in Germany. A vitrectomy is predominantly the course of action in cases where a chronic intraocular leptospiral infection is present. In some cases, the clinical and ophthalmological diagnosis is not conclusive and in these cases a preoperative laboratory test using anterior chamber fluid is indicated. However, false negative results occur when only MAT is used. The aim of this study was to investigate intraocular samples from horses with recurrent uveitis by using ELISA to detect anti-leptospiral antibodies and to compare the results to those of more established laboratory tests, i. e. MAT and PCR. Preliminary investigations indicated a high sensitivity of ELISA in the detection of leptospiral uveitis in horses. 80 eyes of 72 diseased horses were included in the study. The patients' history was taken and a thorough ophthalmologic examination was performed. Intraocular fluids were obtained by diagnostic anterior chamber paracentesis (n = 29) or therapeutic vitrectomy (n = 80) and submitted for MAT, ELISA and PCR to detect leptospiral antibodies and/or DNA. The ELISA allowed the separate determination of the immunoglobulin classes M, G and A. For statistical evaluation Cohen's kappa and the chi-square tests were applied and a p-value of 0.05 or less was considered to be significant. 22 healthy horses (n = 42 eyes) served as controls. 78 % of vitreous samples from horses with ERU and 10 % of healthy controls reacted positively in the MAT, whereas in the ELISA it was 85 % and 0 %, and in the PCR it was 61% and 0 %, respectively. Specific IgA was detected in 84 % of the vitreous samples from horses with ERU, and in 11% of cases, IgA was solely detectable. Differences were highly significant (p<0.001). None of the aqueous samples in the control group contained leptospiral antibodies. Up to 64 % of aqueous humour specimens that were obtained for diagnostic purposes yielded a positive result in the MAT. For the ELISA, it was even 89 %. Anti-leptospiral IgA was the most frequently detected immunoglobulin class. There was a high level of agreement in the MAT and ELISA results. However, ELISA provided a greater number of positive results. The detection of IgA against leptospires proved to be especially sensitive for an intraocular leptospiral infection and, thus, important for the laboratory diagnosis of leptospiral uveitis in horses. Looking at the cost involved in laboratory tests, it would be reasonable to start with an IgA ELISA. Only if there are negative results, additional tests could be performed. In single IgA negative cases there might be positive reactions using MAT, PCR or ELISA for IgG or IgM.

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confidence: 89%

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confidence: 98%

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

Loibl¹,

Gerhards²,

Brem³

et al. 2018

PHK

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“…For some inexplicable reason, recently the calculation of the Goldmann-Witmer coefficient had been revisited as a precondition for the diagnosis of a leptospira-induced uveitis (Gilger 2018, Malalana 2018 as it had been advocated in human ophthalmology for diagnosis of infectious uveitis (Rothova et al 2008). However, an intraocular antibody production in equids had been proved in earlier studies (Wollanke 2002, Wollanke et al 2004) and in addition, PCR and culture results are positive in high percentages , Wollanke et al 2000, Wollanke and Gerhards 2001a, 2001b, Wollanke 2002, Hartskeerl et al 2004, Roczek 2008, Von Borstel et al 2010, Popp et al 2013, Baake et al 2016) and there is ultrastructural evidence of leptospires inside the vitreous of ERU eyes (Niedermaier et al 2006, Brandes et al 2007).…”

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confidence: 99%

“…Detection of antibodies directed against leptospires (Gsell et al 1946, Wood andDavis 1950) and leptospiral DNA in intraocular fluids is not new, but varies considerably between different investigations (Wollanke et al 1998b, Faber et al 2000, Wollanke et al 2001b, Wollanke 2002, Gesell et al 2005, Gilger et al 2008, Loibl 2009, Von Borstel et al 2010, Wiehen 2012, Polle et al 2014, Roth et al 2014, Baake et al 2016, Dorrego-Keiter et al 2016, 2017, Malalana et al 2017. Reasons for this might be the time of sampling (e.g.…”

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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“…Die Operation, Vor-und Nachsorge der ERU-Patienten erfolgte, wie bereits beschrieben (Frühauf et al 1998, von Borstel 2005,Tóth et al 2006, 2010 Auch die suprachoroidale Applikation eines Ciclosporin AImplantats (Gilger et al 2000a, Gilger et al 2000b, Gilger et al 2001, Gilger and Spiess 2006 kann aus Sicht der Autoren bei ERU nicht an die Erfolgsrate der PPV anknüpfen, so dass aufgrund mangelnder therapeutischer Alternativen, einer nach wie vor ungeklärten Ätiopa-thogenese der ERU, regionaler Unterschiede in der Leptospirenprävalenz und der Varianz der MAT-Ergebnisse (Baake et al 2016) …”

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Long-term results of pars plana vitrectomy in relationship to leptospiral antibody detection in vitreous humor in 118 horses with equine recurrent uveitis (ERU)

Keiter¹,

Töth²,

Dikker³

et al. 2017

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Long-term results of pars plana vitrectomy in relationship to leptospiral antibody detection in vitreous humor in 118 horses with equine recurrent uveitis (ERU)In the ongoing discussion regarding aetiopathogenesis and therapeutic approach of equine recurrent uveitis (ERU) it was the aim of the present study to elucidate the relationship of leptospiral antibody detection in vitreous humor and success of pars plana vitrectomy (PPV) in horses with ERU. In a retrospective study of 118 ERU patients of different age, gender and breed undiluted vitreous humor, aseptically taken at the beginning of PPV, was examined by culture. Furthermore vitreous humor and serum were tested for specific antibodies against different serovars of Leptospira interrogans by microscopic agglutination test (MAT). Horses were treated via single port pars plana vitrectomy. The presence or absence of inflammation of the inner eye 8 to 54 months after PPV was evaluated by repeated ocular examinations, as well as by interrogation of referring veterinarians and horse owners. The absence of recurrent inflammation in terms of symptoms of classic uveitis after surgery was considered a success. A total of 55 of 118 horses (46.6 %) had serum antibodies against different serovars of Leptospira (L.) interrogans (≥ 1 : 100). Culture of vitreous humor was positive in 16/118 (13.6 %) patients with ERU. In 49 of 118 horses (41.5 %) and 52 of 134 eyes (38.8 %) leptospiral antibodies were detected in vitreous humor via MAT. L. interrogans serovar Grippotyphosa (48) and L. interrogans serovar Pomona (12) were identified most frequently. A total of 90/118 (76.3 %) horses and 104/134 eyes (77.6 %) remained relapse-free over a period of 8 to 54 months. Forty-one/49 horses (83.7 %) and 44/52 eyes (84.6 %), tested positive for antibodies against L. interrogans via MAT and 49/69 horses (71.0 %) and 60/82 eyes (73.2 %), in which no leptospiral antibodies were detected in vitreous humor, showed no further episodes of ERU (Fishers exact test: P ≥ 0.05). These findings confirm the therapeutic approach to perform a vitrectomy according to the clinical ERU diagnosis irrespective of the leptospiral antibody result.Keywords: horse, equine recurrent uveitis, vitrectomy, leptospira; ophthalmology Zitation: Dorrego Keiter E., Tóth J., Dikker L., Sielhorst J., Schusser G. F. (2017) Long-term results of pars plana vitrectomy in relationship to leptospiral antibody detection in vitreous humor in 118 horses with equine recurrent uveitis

show abstract

Detection of intraocular leptospiral DNA, antibodies and Leptospira spp. in horses with equine recurrent uveitis in different laboratories

Cited by 9 publications

References 48 publications

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

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

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

Long-term results of pars plana vitrectomy in relationship to leptospiral antibody detection in vitreous humor in 118 horses with equine recurrent uveitis (ERU)

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