Evaluation of adenovirus amplified detection of immunochromatographic test using tears including conjunctival exudate in patients with adenoviral keratoconjunctivitis.

BACKGROUND: We evaluated a novel silver amplification immunochromatography test for rapid detection of adenovirus (AdV) antigen equipped with an automated reader system using tears including conjunctival exudate in patients with adenoviral keratoconjunctivitis. METHODS: Two kinds of immunochromatographic (IC) kits, a conventional IC kit for conjunctival scrapings (control kit) and an IC kit using tears including conjunctival exudate collected by pressing a filter paper strip on the conjunctiva (test kit), were tested on 90 patients who attended Migita Eye Clinic with suspected adenoviral conjunctivitis. The results of the test kits were automatically obtained by a specific reader, which was based on silver amplification immunochromatography system, in 15 min. The detection of AdV was confirmed by real-time polymerase chain reaction (PCR) method, and typing was performed by direct sequencing. Comparative dilution assay was carried out with the two kits, using AdV type 3 and type 54 strains. RESULTS: The sensitivity of the control kit and test kit was 89.8% and 98.3%, respectively. The specificity of both kits was 100%. A significant difference in the sensitivities of the two IC kits against PCR positivity was observed (P < 0.01). A significant correlation was found between AdV DNA copy numbers on a logarithmic scale obtained with the two tests (P < 0.01). The sensitivity of the test kit was 32-64-fold higher than that of the control kit without silver amplification for both AdV types. CONCLUSIONS: These results suggest that this novel amplified AdV detection kit using tears including conjunctival exudate is useful, because it decreases patients' discomfort from specimen collection and its sensitivity is significantly higher than that of the conventional IC kit.

Clinical and virological analysis of epidemic keratoconjunctivitis caused by adenovirus type 54 in a regional ophthalmic clinic in Kyushu, Japan.

BACKGROUND: Human adenovirus type 54 (HAdV-54) is a novel type of adenovirus that belongs to species D and has thus far been detected only in Japan in patients with epidemic keratoconjunctivitis (EKC). There was a large nationwide outbreak of HAdV-54 EKC from 2015 to 2016 in Japan. The clinical characteristics of an outbreak of HAdV-54 conjunctivitis treated in a regional ophthalmic clinic in Fukuoka, Japan, in 2016 were analyzed. PATIENTS AND METHODS: A consecutive series of 55 cases diagnosed clinically as EKC confirmed by HAdV-54 detection from conjunctival scrapings by polymerase chain reaction (PCR) method between 17 June 2016 and 29 August 2016 were enrolled. Viral DNA copies were counted by real-time PCR method. The clinical findings were recorded at the first visit to the clinic and evaluated. RESULTS: In the analysis of the relationship between mean clinical score groups and several factors, such as days after onset, sex, HAdV DNA number on a logarithmic scale, and age, most factors did not show a significant difference in clinical score between groups. However, mean clinical score of cases aged <23 years was significantly higher than that of cases aged ≥23 years (P<0.01). The correlation coefficient between DNA copies on a logarithmic scale and clinical score was 0.280, and a significant correlation was observed (P<0.05). Multiple subepithelial corneal infiltrates (MSI) were observed in 24 out of 31 cases (77%). CONCLUSION: These results suggested that the clinical features in the early phase of HAdV-54 keratoconjunctivitis were milder but the rate of MSI observed in the late phase was higher than those in previous epidemics of several HAdV types. The significant difference in clinical severity between age groups might be a virological characteristic of HAdV-54.

Correlation between acute conjunctivitis and Asian dust on ocular surfaces.

The aim of this study was to determine the presence of Asian dust particles (ADP) in patients suffering from conjunctivitis and its correlation with clinical scores for conjunctivitis. Forty-five patients from the Fukuoka area who were newly diagnosed acute conjunctivitis were selected. The degrees of inflammatory reaction, itchy sensation, hyperemia, eye discharge, and foreign body sensation were clinically recorded and scored. Eyes were washed with physiological solution. Solid particles collected from the washing solution were observed using a scanning electron microscope. Of the 45 samples, 44 were positive for the elements silicon (Si) and aluminum (Al), which are components of ambient Asian dust. Higher conjunctivitis scores were found in the subgroup in which the Asian dust/whole particle ratio was greater than average. This is the first apparent report on the correlation between amount of ADP exposure at the ocular surface and severity of ocular symptoms.

Is it possible to detect the presence of adenovirus in conjunctiva before the onset of conjunctivitis?

PURPOSE: It is widely accepted that patients with adenoviral conjunctivitis are contagious approximately as long as 14 days after the onset. However, there is controversy about whether patients with adenoviral conjunctivitis are contagious during the incubation period. We investigated the potential of the eye as a source for spreading infection during the incubation period by analysing the presence of adenovirus (Ad) in contralateral eyes before onset in patients with bilateral adenoviral conjunctivitis using the immunochromatography (IC) test and polymerase chain reaction (PCR). METHODS: Patients with adenoviral conjunctivitis were directed to visit an outpatient clinic at approximately 3-day intervals until onset in the contralateral eye or 2 weeks after onset in the first eye. Conjunctival scrapings obtained from the first infected eye and the contralateral eye before and after onset were tested to detect Ad antigen using the IC test and PCR. RESULTS: Out of the 32 cases, 23 (72%) developed conjunctivitis in the contralateral eye. Among these bilateral cases, 18 (78%) were found positive for Ad by PCR after onset in the contralateral eye. However, no specimen obtained before onset showed a positive result for Ad in both tests. CONCLUSION: Although there is a possibility that eyes are contagious during the incubation period of adenoviral conjunctivitis in some situations, these results suggest that the possibility of the eye acting as a source for spreading infection during the incubation period is very small.

Demographic aspects of allergic ocular diseases and evaluation of new criteria for clinical assessment of ocular allergy.

BACKGROUND: The clinical features of allergic ocular diseases (AOD) are characterized by their wide variety. Clinical evaluation criteria are essential not only for objective assessment in clinical trials, but also for clinical studies on etiological aspects of AOD. However, there have been no internationally established criteria for clinical evaluation and classification of the severity of AOD. We established new criteria and applied them to many cases of AOD, and evaluated their usefulness for clinical purposes. We also studied whether each clinical entity of AOD [allergic conjunctivitis (AC), atopic keratoconjunctivitis (AKC), and vernal keratoconjunctivitis (VKC)] can be distinguished by these criteria. METHODS: A prospective study was carried out to assess the differential diagnosis of 1079 patients with AOD (439 male and 640 female patients). Differential diagnosis of AC, AKC or VKC was made. These diseases were diagnosed and classified based on local and systemic clinical findings. Ten objective ocular clinical findings of conjunctival, limbal and corneal lesions were graded on a 4-point scale, and the total score, with a highest value of 30, was used as the clinical score. RESULTS: Among a total of 1079 cases, seasonal allergic conjunctivitis (SAC), perennial allergic conjunctivitis (PAC), AKC and VKC accounted for 876 (81.2%), 115 (10.6%), 47 (4.4%) and 41 (3.8%) respectively. The mean age in each disease was 52.9, 56.1, 25.7 and 16.6 years respectively. Total clinical score in SAC, PAC, AKC and VKC was 1.54, 2.13, 3.72 and 12.68 respectively. Both mean age and total clinical score in all combinations of two diseases showed significantly different results. CONCLUSIONS: These results suggest that AOD can be classified by our new clinical grading system, and this system is sensitive enough for clinical evaluation of AOD. Re-evaluation of AOD is essential for constructing future strategies for the treatment of AOD, which are consists of various categories of ocular disorders.