Ocular toxoplasmosis: evaluation of lacrimal-specific secretory IgA levels in both patients with active and inactive phases of the disease.

Ocular toxoplasmosis can result in recurrent uveitis. Studies have shown that a correlation between active ocular toxoplasmosis and the presence of anti-Toxoplasma gondii secretory IgA (SIgA) in tears. This study compares anti-T. gondii SIgA levels in patients' tears during the acute and inactive phases of toxoplasmic uveitis. Twenty-nine positive tear specific SIgA for T. gondii patients with acute toxoplasmic uveitis were selected and were followed-up for at least two years, when the anti-T. gondii SIgA tears levels were determined. Specific SIgA for T. gondii was negative in 22 patients (75.86%) and positive in seven patients (24.13%) of whom six (85.7%) were followed over three years. Average SIgA levels during the acute phase are 1.54 and decrease significantly to 0.72 (p = 0.0001) during the inactive phase of disease. Because anti-T. gondii SIgA in the tear is negative in 75.86% of patients after the acute phase of infection, T. gondii SIgA levels may be used as a complementary diagnostic marker for active ocular toxoplasmosis.

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA.

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion) and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9% sensitivity (95% CI = 54.5-74.4), 71.6% specificity (95% CI = 59.8-81.2), a positive predictive value of 72% (95% CI = 60.3-81.5) and a negative predictive value of 65.4% (95% CI = 54.0-75.4). sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05). The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.

Intense Pulsed Light Therapy for Patients with Meibomian Gland Dysfunction and Ocular Demodex Infestation.

To observe the clinical changes of meibomian gland dysfunctipn (MGD) and ocular Demodex infestation after intense pulsed light (IPL) treatment to further examine the mechanism of IPL treating patients with MGD and ocular Demodex infestation. The medical records of 25 patients (49 eyes) with MGD treated with IPL, were retrospectively examined to determine outcomes. Associated ocular-surface parameters (ocular surface disease index, OSDI; lipid layer thickness, LLT; noninvasive first breakup time, NIF-BUT; noninvasive average breakup time, NIAvg-BUT; tear film breakup area, TBUA; Schirmer I Test, SIT; corneal fluorescein staining, CFS), eyelid margin abnormalities, meibum quality and expressibility, MG morphological parameters (macrostructure and microstructure), and the number of Demodex infestation were examined before and after treatment. The MG microstructure and the Demodex infestation were examined via in vivo confocal microscopy (IVCM). The results showed that there were statistically significant differences in associated ocular-surface parameters (all P<0.05) before and after IPL treatment, except SIT (P=0.065). Eyelid margin abnormalities, meibum quality and expressibility obviously improved in upper and lower eyelid after IPL treatment (all P<0.0001). MG macrostructure (MG dropouts) decreased in upper (P=0.002) and lower eyelid (P=0.001) after IPL treatment. The nine parameters of MG microstructure in upper and lower eyelid all distinctly improved after IPL treatment (all P<0.0001). The mean number of Demodex mites on the upper lid margin (6.59±7.16 to 3.12±3.81/9 eyelashes) and lower lid margin (2.55±2.11 to 1.29±1.53/9 eyelashes) significantly reduced after IPL treatment (all P<0.0001). The Demodex eradication rate was 20% (8/40) in upper lid margin and 34.15% (14/41) in lower lid margin. These findings indicate that IPL shows great therapeutic potential for patients of MGD and ocular Demodex infestation.

[Development of an antigen detection dot blot assay for the diagnosis of human onchocerciasis based on the biotin-avidin binding system]. / Développement d'un test dot blot de détection d'antigène, basé sur le système de fixation biotine- avidine, pour le diagnostic de l'onchocercose humaine.

The development of a highly sensitive and specific diagnostic test is of urgent need for the field assessment of human onchocerciasis and for monitoring the success of control programs. We report here the development and evaluation of a Dot blot Immunobinding Assay (DIA-BA) based on the biotin-avidin binding system, for the detection of O. volvulus specific antigens in body fluids. Specific antibodies were produced by immunizing rabbits with the O. volvulus recombinant antigen Oncho-C71 and labelled with biotin. The biotinylated probes were then used to detect O. volvulus specific antigens initially blotted onto a nitrocellulose membrane. The smallest amount of blotted antigens detectable by the new test is 0.5ng, 1ng, 1ng and 2ng respectively in urine, dermal fluid, tears and serum samples. Out of 456 onchocerciasis endemic subjects examined, 98.4%, 96.5%, 90.8% and 75.0% were positive by the DIA-BA test on urine, dermal fluid, tears and serum respectively The test was most sensitive (100%) when used on urine and least (54.76%) when used on serum from skin snip positive subjects. The specificity of the test, determined amongst non-exposed individuals, was 100% on all but for dermal fluid samples (97.5%). Also, the color intensities on the blot were observed to positively correlate (r = 0.8 on urine) with the skin microfilaria loads on the individuals. We conclude that DIA-BA test could be very useful for mass diagnosis of prepatent, of low and high level infections due to O. volvulus.

Polymerase chain reaction analysis of corneal epithelial and tear samples in the diagnosis of Acanthamoeba keratitis.

PURPOSE: Acanthamoeba is an uncommon cause of corneal infection in which the best visual outcome follows prompt diagnosis and a long course of appropriate antimicrobial therapy. Because conventional detection techniques for Acanthamoeba have certain limitations, we investigated the ability of the polymerase chain reaction (PCR) to confirm the clinical diagnosis of Acanthamoeba keratitis, with the ultimate aim of achieving early diagnosis. METHODS: Using two different pairs of primers, PCR was performed on representative cultured Acanthamoeba isolates to confirm the assay's ability to amplify Acanthamoeba DNA from a wide range of acanthamoebae. Subsequently, corneal epithelial samples from 19 patients and tear samples from 12 patients with Acanthamoeba keratitis were analyzed by PCR for the presence of Acanthamoeba DNA. RESULTS: Acanthamoeba DNA was amplified by PCR from 16 (84%) of 19 corneal epithelial samples, whereas Acanthamoeba was cultured from 10 samples (53%), all of which were PCR positive. Tear samples from 8 (66%) of 12 patients were positive on PCR testing, and one tear sample was PCR positive, whereas the corresponding epithelial biopsy had yielded a negative PCR result. Samples from culture-positive patients were positive on PCR testing more frequently than those from culture-negative patients (10/10 culture-positive corneal epithelial and 5/7 [71%] culture-positive initial tear samples versus 6/9 [66%] culture-negative corneal epithelial and 2/5 [40%] culture-negative tear samples). All control epithelial (n = 15) and tear (n = 15) samples yielded negative results. CONCLUSIONS: PCR was a more sensitive diagnostic test than a culture for Acanthamoeba keratitis, and the use of two different primers achieved better sensitivity than a single set. A PCR of a tear sample also may be a useful complementary test and, in combination with PCR of epithelial samples, would prove particularly helpful in confirming the clinical diagnosis in culture-negative cases.

Association between Pelodera strongyloides (Nematoda: Rhabditidae) and wood mice, Apodemus sylvaticus.

Pelodera strongyloides larvae were found in the conjunctival sacs of Apodemus sylvaticus and Clethrionomys glareolus and in hair follicles of A. sylvaticus. Those in the eyes were 3rd-stage larvae while most of those in the dermis were early 4th stages. There was no evidence of pathology and sections of the skin showed that the nematodes of freshly killed mice and moulted once to the adult stage in 1-2 days at 15 degrees C. The stimulus to resume development appeared to be a reduction in temperature rather than presence of bacterial food or reduction in osmotic pressure. Countless generations ensued on nutrient agar provided the nematodes were periodically sub-cultured onto fresh agar. Nematodes from the eyes died in culture. Dauerlarvae of P. strongyloides were produced in exhausted cultures. They resumed development on fresh agar but not in distilled water. About half could be induced to exsheath by a temperature similar to that of mouse skin. A. sylvaticus probably accumulates nematode larvae in the dermis during its life and the nematodes resume development when the host dies.

Biology of Toxoplasma gondii, its survival in body tissues and liquids, risks for the pregnant woman.

After briefly outlining the biology of Toxoplasma gondii the authors discuss the resistance of the parasite in body liquids and tissues under certain environmental conditions. Parasite resistance and its consequent risk of human infection, particularly for the pregnant woman, is emphasized. Toxoplasma oocysts, the sexual expression of the parasite, and the cyst facilitate the diffusion of Toxoplasma, as they are much more resistant than the trophozoite.

Anti-Toxoplasma gondii secretory IgA in tears of patients with ocular toxoplasmosis: immunodiagnostic validation by ELISA

Toxoplasma gondii causes posterior uveitis and the specific diagnosis is based on clinical criteria. The presence of anti-T. gondii secretory IgA (sIgA) antibodies in patients' tears has been reported and an association was found between ocular toxoplasmosis and the anti-T. gondii sIgA isotype in Brazilian patients. The purpose of this study was to provide an objective validation of the published ELISA test for determining the presence of anti-T. gondii sIgA in the tears of individuals with ocular toxoplasmosis. Tears from 156 patients with active posterior uveitis were analysed; 82 of them presented characteristics of ocular toxoplasmosis (standard lesion) and 74 patients presented uveitis due to other aetiologies. Cases of active posterior uveitis were considered standard when a new inflammatory focus satellite to old retinochoroidal scars was observed. The determination of anti-T. gondii sIgA was made using an ELISA test with crude tachyzoite antigenic extracts. Tears were collected without previous stimulation. Detection of sIgA showed 65.9 percent sensitivity (95 percent CI = 54.5-74.4), 71.6 percent specificity (95 percent CI = 59.8-81.2), a positive predictive value of 72 percent (95 percent CI = 60.3-81.5) and a negative predictive value of 65.4 percent (95 percent CI = 54.0-75.4). sIgA reactivity was higher in the tears of patients with active posterior uveitis due to T. gondii (p < 0.05). The test is useful for differentiating active posterior uveitis due to toxoplasmosis from uveitis caused by other diseases.