Rapid Pathogen Screening Research Articles

Three birds with one stone: A nano zeolitic imidazolate framework-luciferase-antimicrobial peptide biocomposite-based biosensing platform for improving enzyme stability, detection sensitivity, and sterilization effect

Adenosine triphosphate-driven bioluminescence (ATP-BL) biosensors were widely employed for on-site screening of bacteria. However, unstable luciferase with strict storage conditions limited its field detection. Moreover, the traditional ATP-BL biosensors lacked the sterilization effect. In this study, a robust and multifunctional zeolitic imidazolate framework-90 decorated with luciferase and polyethylene glycol–antimicrobial peptides (ZIF-90@Luc-AMP) was prepared. It could not only enhance the activity and stability of luciferase at ambient temperatures (≥1 month) but also sensitively detect target bacteria via the bioluminescence (BL) method and kill them (three birds with one stone). During the detection process, the target pathogen Escherichia coli O157:H7 (E. coli O157:H7) was specifically captured on a phage-modified stir bar and formed a sandwich complex with ZIF-90@Luc-AMP. With the synergistic bacteriolysis of ZIF-90@Luc-AMP and the phage, the captured E. coli O157:H7 was decomposed and emitted adenosine triphosphate (ATP), achieving the sterilization effect. Meanwhile, the introduced luciferase catalyzed ATP to produce a BL signal, which was proportional to the concentration of E. coli O157:H7. Combining ZIF-90@Luc-AMP and a phage-labeled stir bar for target recognition and enrichment, the entire analytical process could be easily manipulated within 30 min with a low limit of detection (20 CFU/mL) through a portable ATP bioluminescent meter. The multifunctional ZIF-90@Luc-AMP biosensor with high stability, specificity, and sensitivity was also appropriate for rapid pathogen screening and antibacterial application in the wild.

Molecular analysis of human adenoviral keratoconjunctivitis cases: Results of a 2-year survey

Objective: This study aimed to determine the adenovirus genotypes and their epidemiological features between January 2018 and
 November 2019, in Istanbul, Turkey.
 Material and Methods: Conjunctival swab samples were obtained from patients who were clinically diagnosed with keratoconjunctivitis.
 Samples were screened with an Adeno Detector kit (Rapid Pathogen Screening, RPS Inc., South Williamsport, PA). Nucleic acid
 extraction and amplification were performed with the ADENOVIRUS ELITe MGB® kit in the ELITe In Genius instrument (Elitech
 Group, Torino, Italy). For subtyping of the strains, sequencing primers targeted the ‘Hypervariable Region 7’ (HVR-7) of the hexon
 gene were used. DNA sequence analysis (n:72) was performed with ABI PRISM® 3100 Genetic Analyzer (Applied Biosystems, USA),
 and subtyping was done by BLAST analysis.
 Results: The median viral load in the samples (n: 77) was 7 log10 copies/mL (IQR: 4.5-7.4 log10 copies/mL). The clinical finding score
 was found to be significantly higher in the high viral load group (Adenovirus DNA≥6 Log 10 copies/mL) than in the low viral load
 group (Adenovirus DNA

Development of a diagnostic assay by three-tube multiplex real-time PCR for simultaneous detection of nine microorganisms causing acute respiratory infections

Acute respiratory infections are widespread in vulnerable populations of all ages and are characterized by a variety of symptoms. The underlying infection can be caused by a multitude of microorganisms, including viruses and bacteria. Early detection of respiratory infections through rapid pathogen screening is vital in averting infectious respiratory disease epidemics. This study utilized a multiplex real-time PCR system to develop a three-tube reverse transcription-PCR (RT-PCR) assay, enabling simultaneously detect nine respiratory pathogens, including: influenza A and B, adenovirus, respiratory syncytial virus (RSV), Streptococcus pneumoniae, Legionella pneumophila, Haemophilus influenzae, Chlamydia pneumoniae, and Mycoplasma pneumoniae. This technique utilizes a one-step assay, with specifically designed TaqMan primer–probe sets combined in the same tube. This assay provided rapid and simplified detection of the nine prevalent pathogens, as well as increased sensitivity and reduced cross-contamination. This assay was evaluated using 25 related viral/bacterial strains as positive references, the other 25 irrelevant strains as negative controls, and clinical specimens from 179 patients. All positive strains were detected with no amplification of the non-target microorganism mixtures and the assay’s detection limits ranged between 250–500 copies/ml (1.25–2.5 copies/reaction). A total of 167 (93.3%) samples tested positive for at least one of the pathogens identified; 109 of these samples were from patients confirmed to have RSV infections. The diagnostic accuracy of our assay was further confirmed by matching results from classical direct immunofluorescence assay and nucleotide sequencing. These data demonstrate the innovative multiplex real-time PCR assay as a promising alternative to the current approaches used for early screening of acute respiratory infections.

Changes in the Matrix Metalloproteinase 9 Point-of-Care Test Positivity According to MMP-9 Concentration and Loading Volume.

To measure changes in the matrix metalloproteinase 9 (MMP-9) point-of-care test, InflammaDry (Rapid Pathogen Screening, Inc, Sarasota, FL) positivity, based on ocular surface MMP-9 concentrations and loading volume. Two different MMP-9 products, preform and active, were analyzed using the InflammaDry test, detecting MMP-9 levels of more than 40 ng/mL of both preform and active MMP-9. Preform MMP-9 (Natural human MMP-9 protein; Abcam, Cambridge, UK) was analyzed at different concentrations (50, 100, 500, 1000, and 1500 ng/mL) and loading volumes (5, 10, and 20 μL). Active MMP-9 (Human MMP-9 protein; Novus Biologicals, Littleton, CO) was also analyzed using the InflammaDry test at different concentrations (50 and 100 ng/mL) and loading volumes (10, 20, and 40 μL). Natural human MMP-9 protein (preform) of 50, 100, and 500 ng/mL exhibited negative results for every loading volume. At 1000 ng/mL, the 20 μL volume was positive, whereas the 5 and 10 μL volumes were negative. At 1500 ng/mL, all loading volumes were positive, but the density of positive bands varied depending on the loading volume; larger loading volumes had higher band density. Human MMP-9 protein (active) of 50 ng/mL was negative for every loading volume. In 100 ng/mL, the 20 and 40 μL volumes showed positive results with similar positive band densities. The InflammaDry test had a different detection range depending on MMP-9 formulas; higher concentrations of preform MMP-9 protein were needed to yield positive results. In addition, InflammaDry positivity varied based on the loading volumes. Clinicians should be aware of the possibility of false negatives with low tear volumes despite elevated MMP-9 concentrations.

Corneal biomechanical alterations in patients with chronic ocular Graft Versus-Host Disease.

PurposeTo compare corneal biomechanics between patients with ocular graft versus-host disease (oGVHD) and healthy subjects (controls), and to further correlate these values with ocular and hematological characteristics.Materials and methodsThe following procedures were performed in oGVHD patients and controls: Schirmer test (ST), break-up time (BUT), corneal and conjunctival staining, tear matrix metalloproteinase-9 (MMP-9) assay (InflammaDry test, Rapid Pathogen Screening, Inc, Sarasota, FL). Corneal biomechanics were calculated by using ocular response analyzer (ORA, Reichert Instruments, Depew, New York, USA). The Mann-Whitney U test was used to compare continuous variables between oGVHD patients and controls. Correlations of corneal biomechanics with ocular and hematological parameters were examined using Spearman's correlation.ResultsA total of 45 oGVHD patients (mean age ± SD, 51.5 ± 7.1 years) and 34 controls (47.8 ± 6.1 years) were included. Patients with oGVHD showed significantly lower values of corneal hysteresis (CH) and corneal resistance factor (CRF) compared to controls (respectively, 9.4 ± 1.8 mmHg vs 11.6 ± 1.6 and 9.7 ± 1.4 mmHg vs 12.3 ± 1.3; always p<0.001). Twenty-nine of the oGVHD eyes (64.4%) were strong-positive for MMP-9, while 16 (35.6%) were weak-positive. Conversely, only 4 of the control eyes (11.8%) were weak-positive for MMP-9. In patients with oGVHD, CH was significantly correlated with corneal staining (Rs = -0.316, p = 0.035), conjunctival staining (Rs = -0.437, p = 0.003), ST (Rs = 0.390, p = 0.008), BUT (Rs = 0.423, p = 0.004), oGVHD severity grade (Rs = -0.383, p = 0.009), and MMP-9 positivity grade (Rs = -0.429, p = 0.003), while CRF was correlated only with corneal staining (Rs = -0.317, p = 0.034).ConclusionsCorneal biomechanics are reduced in patients with oGVHD, and CH is negatively correlated with disease severity grade and MMP-9 tear levels.

2023. A Prospective, Multi-Center U.S. Clinical Trial to Determine Accuracy of FebriDx Point-of-Care Testing for Acute Upper Respiratory Infections with and Without a Confirmed Fever

BackgroundFebriDx is a 10-minute disposable point-of-care test designed to identify clinically significant systemic host immune responses and aid in the differentiation of viral and bacterial respiratory infection by simultaneously detecting C-reactive protein (CRP) and myxovirus resistance protein A (MxA) from a fingerstick blood sample.MethodsA prospective, multicenter, cross-sectional study of primarily adults with acute upper respiratory tract infections (URIs), with and without a confirmed fever at the time of enrollment, was performed to evaluate the diagnostic accuracy of FebriDx to identify clinically significant bacterial infection with host response and acute pathogenic viral infection. URI was defined as rhinosinusitis, pharyngitis, nonspecific URI, and bronchitis and the reference method consisted of an algorithm that included throat bacterial cell culture, respiratory PCR panels for viral and atypical pathogens, procalcitonin, white blood cell count, and bandemia. The algorithm also utilized the Centor criteria and allowed for physician over-ride.ResultsAmong 220 patients enrolled, 100% reported fever ≥100.5 within the last 72 hours while 55% (121/220) had a confirmed fever at the time of enrollment. Of the total enrolled patients, 15% (34/220) were classified as bacterial, 56% (124/220) were classified as viral, and 28% (62/220) negative by the reference standard. Sample Size (n) Confirmation of Fever Diagnosis Sensitivit % and [95% CI] Specificity % and [95% CI] Positive Predictive Value (PPV)% and [95% CI] Negative Predictive Value (NPV)% and [95% CI] 220Reported within last3 daysBacterial 85 (29/34) [69-95] 93 (183/196) [89-96] 69 (29/42 ) [56-79] 97 (183/188) [94-99]Viral 90 (111/124) [83-94] 76 (73/96) [66-84] 83 (111/134) [77-87] 85 (73/86) [77-90]121Exhibited on enrollmentBacterial 95 (19/20) [77-100] 94 (95/101) [88-98] 76 (19/25) [59-87] 99 95/96) [93-100]Viral 90 (72/80) [81-96] 78 (32/41) [62-89] 89 (72/81) [82-93] 80 (32/40) [67-89]ConclusionWhen comparing clinical accuracy of diagnostic tests, performance values should be determined in febrile patients. FebriDx’s 97–99% NPV may help to identify clinically significant bacterial URI’s and supports outpatient antibiotic decisions.Disclosures R. Sambursky, Rapid Pathogen Screening, Inc.: Board Member, Employee and Shareholder, Salary.

Randomized, Controlled, Phase 2 Trial of Povidone-Iodine/Dexamethasone Ophthalmic Suspension for Treatment of Adenoviral Conjunctivitis

To evaluate the efficacy/safety of an ophthalmic suspension of povidone-iodine (PVP-I) 0.6% and dexamethasone 0.1% in patients with acute adenoviral conjunctivitis. Multicenter, randomized, vehicle-controlled, double-masked trial. Adults with a positive Rapid Pathogen Screening Adeno-Detector Plus test were randomized 1:1:1 to PVP-I 0.6%/dexamethasone 0.1%, PVP-I 0.6%, or vehicle, bilaterally 4 times daily for 5days (days 1-5). Patients were evaluated on days 3, 6, and 12 (+1-day window). Efficacy measures included clinical resolution and adenoviral eradication. Overall, 144 patients were included in the efficacy analysis (PVP-I/dexamethasone, n= 48; PVP-I, n= 50; vehicle, n= 46). The proportion of patients with clinical resolution (primary study eye with last observation carried forward [LOCF]) at the day 6 visit was higher with PVP-I/dexamethasone (31.3%) than with vehicle (10.9%; P= .0158) and PVP-I (18.0%; P= nonsignificant). The proportion with adenoviral eradication (primary study eye with LOCF) was higher with PVP-I/dexamethasone than with vehicle at the day 3 (35.4% vs 8.7%; P= .0019) and day 6 (79.2% vs 56.5%; P= .0186) visits and vs PVP-I (day 3 visit, 32.0%; day 6 visit, 62.0%; each P= nonsignificant). Treatment-emergent adverse events (AEs) occurred in 69.0% (vehicle), 62.7% (PVP-I), and 53.4% (PVP-I/dexamethasone) of patients in the safety dataset. Discontinuation owing to AEs occurred in 37 patients (vehicle, n= 16; PVP-I, n= 12; PVP-I/dexamethasone, n= 9). PVP-I/dexamethasone appeared safe and well tolerated, and significantly improved clinical resolution and adenoviral eradication in patients with acute adenoviral conjunctivitis.

Role of the Adenoplus test in refractory, recurrent and clinically undiagnosed conjunctivitis

BackgroundIn diagnosing adenoviral conjunctivitis, polymerase chain reaction (PCR) is widely adopted as a diagnostic tool. A new antigen-based immunoassay test (AdenoPlus; Rapid Pathogen Screening Inc, Sarasota, Fla.) is commercially available as an alternative diagnostic test. To date, evidence around the role of this test in the clinical setting has been limited and contradictory. ObjectiveTo determine the sensitivity and specificity of the AdenoPlus test relative to PCR in detecting the presence of adenovirus in patients with conjunctivitis that is recurrent, refractory to treatment, or clinically of unknown etiology. MethodsA prospective study of 27 patients presenting to an acute eye clinic with conjunctivitis that is recurrent (Group A), refractory to treatment (Group B), or clinically of unknown etiology (Group C). All patients underwent the AdenoPlus test and PCR analysis. Sensitivity and specificity were calculated for AdenoPlus using PCR as a reference standard. ResultsOf 27 patients, 7% were in Group A, 19% in Group B, and 74% in Group C. Relative to PCR, the AdenoPlus test demonstrated a sensitivity of 33.3% (95% CI 4% to 78%) and specificity was 95.2% (95% CI 76% to 100%). Positive predictive value was 66.7% (95% CI 9% to 99%); negative predictive value was 83.3% (95% CI 63% to 95%). ConclusionsDue to its high specificity, AdenoPlus may be a good diagnostic test, although further study is indicated. Due to its low sensitivity, however, the test should not be used as a screening tool in patients presenting with these features of conjunctivitis.

Matrix Metalloproteinase 9 Testing in Dry Eye Disease Using a Commercially Available Point-of-Care Immunoassay

To measure matrix metalloproteinase 9 (MMP-9) in the tear film of patients with dry eye disease (DED) compared with controls and to correlate clinical findings. In a prospective study, 101 patients and controls underwent MMP-9 testing of the tear film. Thereafter, they were evaluated for symptoms and signs of DED. Included patients were those who showed 3 of the following 4 dry eye criteria: ocular surface disease index (OSDI) score of more than 12, tear film break-up time (TBUT) of 10 seconds or less, Schirmer test results without anesthesia of less than 10 mm/5 minutes, and corneal staining results of 1 or more. Fifty-four healthy eyes and 47 eyes fulfilling diagnostic criteria for DED of various levels of severity were included in this study. The tear film was analyzed for MMP-9 by a commercially available test (InflammaDry; Rapid Pathogen Screening, Inc, Sarasota, FL) detecting MMP-9 levels of more than 40 ng/ml. Symptoms and signs of DED were evaluated using the OSDI questionnaire, TBUT, conjunctival and corneal staining, Schirmer test results without anesthesia, and meibomian gland examination. These findings were correlated to results of the MMP-9 test in tears. Positive MMP-9 results in tears. In 19 of 47 patients confirmed with dry eye (40.4%) and in 3 of 54 controls (5.6%), the MMP-9 results were positive. This difference was statistically significant (P < 0.001). Thus, the MMP-9 results indicated a clinically significant inflammation in 40% of dry eye patients. Positive results correlated well with subjective symptoms of DED evaluated by OSDI (P= 0.001), TBUT of less than 5 seconds (P < 0.013), Schirmer test results (P < 0.001), conjunctival staining (P < 0.001), and corneal staining (P= 0.007). Moreover, MMP-9 results correlated with the number of obstructed meibomian ducts (P= 0.005) and a pathologic meibomian gland secretion (P= 0.001). The MMP-9 results were increased significantly in women (P < 0.001) and in patients with autoimmune disease (P= 0.005), especially Sjögren's syndrome (P= 0.001) and thyroid disease (P= 0.012). Matrix metalloproteinase 9 testing in DED is a valuable new diagnostic tool. It correlated well with other dry eye tests and identified the presence of ocular surface inflammation in 40% of confirmed dry eyepatients. It may be especially helpful to identify patients with ocular surface inflammation and autoimmune disease and may facilitate the decision to institute anti-inflammatory treatment in these patients.

Droplet Microfluidic Chip Based Nucleic Acid Amplification and Real-Time Detection of Influenza Viruses

Miniaturized bio-diagnostic devices have the potential to allow for rapid pathogen screening in clinical patient samples, as a low cost and portable alternative to conventional bench-top equipment. Miniaturization of key bio-diagnostic techniques, such as: nucleic acid detection and quantification, polymerase chain reaction (PCR), DNA fingerprinting, enzyme linked immunosorbent assay (ELISA), results in substantial reduction of reaction volumes (expensive samples/reagents) and shorter reaction times. Droplet microfluidics (DMF) is one of several miniaturized bio-sample handling techniques available for manipulating clinical samples and reagents in microliter (10−6 L) to picoliter (10−12 L) volume regime. Electro-actuation of sample and reagent in the form of droplets in the aforementioned volume regime, using dielectrophoresis (DEP) and/or Electrowetting (EW) are achieved by means of patterned, insulated metal electrodes on one or more substrates. In this work, we have utilized electro-actuation based DMF technology, integrated with suitably tailored resistive micro-heaters and temperature sensors, to achieve chip based real-time, quantitative PCR (qRT-PCR). This qRT-PCR micro-device was utilized to detect and quantify the presence of influenza A and C virus nucleic acids, using in-vitro synthesized viral RNA segments. The experimental analysis of the DMF micro-device confirms its capabilities in qRT-PCR based detection and quantification of pathogen samples, with accuracy levels comparable to established commercial bench-top equipment (PCR efficiency ∼95%). The limit of detection (LOD) of the chip based qRT-PCR technique was estimated to be ∼5 copies of template RNA per PCR reaction.

Immunoassay für Matrix-Metalloproteinase-9 in der Tränenflüssigkeit bei Patienten mit Pseudoexfoliationssyndrom - eine Pilotstudie

Matrix-metalloproteinases (MMPs) are proteolytic enzymes released by irritated epithelial cells of the ocular surface. It has been established that the subtype MMP-9 can serve as an inflammatory marker within the tear film. MMP-9 is also attributed to have an effect on the PEX-glaucoma development. Recently, a rapid immunoassay for detection of MMP-9 in the tear film was developed to estimate inflammatory extent during dry eye disease. The aim of this study was to analyse the MMP-9 concentration in tear film in PEX-syndrome. In addition, an assessment of the feasibility, reliability and readability of the test was done. We randomly selected 10 patients with PEX-syndrome and 10 healthy control patients and measured tear film MMP-9 of one eye with the RPS InflammaDry Detector™ (Rapid Pathogen Screening Inc., USA). We detected increased levels of MMP-9 in tear film in PEX-syndrome. 80 % of the PEX-patients and 20 % of the controls showed a positive test result (>or= 40 ng/mL MMP-9) indicating a test specificity and sensitivity of 80 %. This corresponds approximately to the published values for the dry eye (sensitivity 87 %, specificity: 92 %). The performance of the test is simple. The patients tolerated the inclusion of the test strips well. However, it is difficult to estimate whether enough tear film was used and in many cases, the intensity of the "indicator line" was weak. The rapid MMP-9-immunoassay is a novel, meaningful approach for the detection of inflammatory activity of the ocular surface. We have shown an up-regulation of the non-specific inflammatory marker MMP-9 in tear film in PEX-syndrome and suggest an association with a tear film disorder. However, an improvement in the estimation of the amount of collected tears and readability is desirable.

The Practical Detection of MMP-9 Diagnoses Ocular Surface Disease and May Help Prevent Its Complications

To evaluate the importance and practicality of testing for matrix metalloproteinase 9 (MMP-9) in dry eye and ocular surface disease. This enzyme, which can cause tissue damage, seems also to be the most reliable diagnostic indicator of ocular surface disease. Enzyme-linked immunosorbent assay, polymerase chain reaction, diffusion, and InflammaDry, a new rapid immunoassay by RPS (Rapid Pathogen Screening Inc). MMP-9 measurement is sensitive and accurate for diagnosing dry eye and ocular surface disease and compares favorably in both sensitivity and specificity against the existing methods of dry eye diagnosis. Abnormal elevations of MMP-9 may predict post-laser in situ keratomileusis complications and refractive complications such as epithelial ingrowth and corneal ulceration. The presence of elevated MMP-9 on the ocular surface will identify those patients who should receive antiinflammatory therapy, such as cyclosporine, and may predict those patients who will respond to this therapy. A rapid in-office test that is sensitive for identifying inflammatory dry eye and ocular surface disease may facilitate better preoperative management of the ocular surface. Optimization of the ocular surface perioperatively would be expected to reduce complications from laser in situ keratomileusis and other surgeries that often make the underlying disease worse. This test may also indicate the need for antiinflammatory therapies, such as cyclosporine or steroids, and also may predict those patients who are more likely to respond.

Next‐generation sequencing: ready for the clinics?

Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing and increasing the throughput. Now, the focus is on using NGS technology for diagnostics and therapeutics. In this review, we discuss the possible clinical applications of NGS and the potential of some of the current systems to transition to the clinic. Clinical use of NGS technologies will enable the identification of causative mutations for rare genetic disorders through whole-genome or targeted genome resequencing, rapid pathogen screening and cancer diagnosis along with the identification of appropriate therapy. Routine clinical use of NGS technologies is appealing, but mandates high accuracy, simple assays, small inexpensive instruments, flexible throughput, short run times and most importantly, easy data analysis as well as interpretation. A number of NGS systems launched recently have least some of these characteristics, namely, small instruments, flexible throughput and short run time, but still face a few challenges. Moreover, simplified data analysis tools will need to be developed to minimize the requirement of sophisticated bioinformatics support in clinics. In summary, for successful transition of NGS to clinic, a sustained collaboration between research labs, clinical practitioners and vendors offering sequencing based genetic tests is required.

Adenovirus advances: new diagnostic and therapeutic options

Adenoviral infection is common, can be severe, and may cause significant morbidity. Ophthalmologists and optometrists are often guilty of spreading adenovirus because it is highly contagious and has 53 serotypes with variable morphology. Adenovirus is often difficult to diagnose based on clinical appearance and, in the early stages, is associated with a red eye or superficial keratitis common to herpes and other infections. This difficulty results in the indiscriminate use of antibiotics, which are expensive and of no established value in treating a viral infection. The difficulty of accurate diagnosis also makes the use of newer proposed treatments less valuable and even potentially hazardous. New diagnostic tests such as the Rapid Pathogen Screening (RPS) Adeno Detector that are practical, rapid, and inexpensive to use in the office may obviate these problems.

A combination povidone-iodine 0.4%/dexamethasone 0.1% ophthalmic suspension in the treatment of adenoviral conjunctivitis

The objective of this pilot study was to determine the preliminary efficacy of a novel ophthalmic suspension containing povidone-iodine 0.4% and dexamethasone 0.1% in the treatment of adenoviral conjunctivitis. A prospective, open-label, single-armed, phase II clinical trial in humans. Eligible patients with the clinical signs and symptoms of acute conjunctivitis who tested positive for adenoviral antigen by Rapid Pathogen Screening (RPS) Adeno Detector were enrolled in a single treatment arm consisting of a combination povidone-iodine 0.4%/dexamethasone 0.1% sterile ophthalmic suspension given four times daily for a minimum of 5 days. RPS Adeno Detector testing was performed at baseline and at each follow-up visit along with ocular fluid sampling by conjunctival swabs. Subsequent analysis performed on all swabs included both adenoviral titer by quantitative polymerase chain reaction (qPCR) and cell culture with confirmatory immunofluorescence (CC-IFA). The primary endpoint was clinical resolution of conjunctival injection and discharge. Secondary measures included reduction of qPCR titers and eradication of infectious virus as determined by CC-IFA. A total of nine eyes of six patients with clinical signs and symptoms of acute viral conjunctivitis and a positive RPS Adeno Detector test result were enrolled in the study. In eight/nine eyes enrolled in the study, clinical resolution was observed by day 3 or day 4. In six/six eyes with detectable adenovirus by qPCR, significant reduction in viral titer was seen by day 3, day 4, or day 5. In five/six eyes with infectious virus confirmed by CC-IFA at enrollment, elimination of infectivity was achieved by day 4 or day 5. One patient was lost to followup. An ophthalmic suspension containing povidone-iodine 0.4% and dexamethasone 0.1% may be a useful agent in the treatment of acute RPS Adeno Detector-positive conjunctivitis. A further placebo-controlled study with a larger number of patients is warranted.

The RPS Adeno Detector for Diagnosing Adenoviral Conjunctivitis

To compare the sensitivity, specificity, and accuracy of the RPS Adeno Detector (Rapid Pathogen Screening Inc., South Williamsport, PA) against both viral cell culture with confirmatory immunofluorescence staining (CC-IFA) and the polymerase chain reaction (PCR) for diagnosing adenoviral conjunctivitis. Prospective, nonrandomized, masked, multicenter clinical trial. One hundred eighty-six consecutive patients from 5 clinical centers seeking treatment within 1 week of developing a red eye and thought to have acute conjunctivitis. The RPS Adeno Detector is a 10-minute in-office lateral flow immunoassay. Patients were tested with the RPS Adeno Detector, CC-IFA, and PCR to detect the presence of adenovirus. The sensitivity, specificity, and accuracy of the RPS Adeno Detector were assessed for identifying cases of adenoviral conjunctivitis. Compared with CC-IFA, the RPS Adeno Detector was 88% sensitive and 91% specific at detecting adenoviral conjunctivitis. Using PCR as a reference method, the sensitivity of the RPS Adeno Detector increased to 89% and the specificity increased to 94%. Compared with PCR, CC-IFA was found to be 91% as sensitive and 100% as specific. The RPS Adeno Detector demonstrated sufficient sensitivity and specificity to be used in the physician's office for the detection of adenoviral conjunctivitis.

Mucin and Leukocyte Interactions In Vitro

To measure matrix metalloproteinase 9 (MMP-9) in the tear film of patients with dry eye disease (DED) compared with controls and to correlate clinical findings.In a prospective study, 101 patients and controls underwent MMP-9 testing of the tear film. Thereafter, they were evaluated for symptoms and signs of DED.Included patients were those who showed 3 of the following 4 dry eye criteria: ocular surface disease index (OSDI) score of more than 12, tear film break-up time (TBUT) of 10 seconds or less, Schirmer test results without anesthesia of less than 10 mm/5 minutes, and corneal staining results of 1 or more. Fifty-four healthy eyes and 47 eyes fulfilling diagnostic criteria for DED of various levels of severity were included in this study.The tear film was analyzed for MMP-9 by a commercially available test (InflammaDry; Rapid Pathogen Screening, Inc, Sarasota, FL) detecting MMP-9 levels of more than 40 ng/ml. Symptoms and signs of DED were evaluated using the OSDI questionnaire, TBUT, conjunctival and corneal staining, Schirmer test results without anesthesia, and meibomian gland examination. These findings were correlated to results of the MMP-9 test in tears.Positive MMP-9 results in tears.In 19 of 47 patients confirmed with dry eye (40.4%) and in 3 of 54 controls (5.6%), the MMP-9 results were positive. This difference was statistically significant (P < 0.001). Thus, the MMP-9 results indicated a clinically significant inflammation in 40% of dry eye patients. Positive results correlated well with subjective symptoms of DED evaluated by OSDI (P = 0.001), TBUT of less than 5 seconds (P < 0.013), Schirmer test results (P < 0.001), conjunctival staining (P < 0.001), and corneal staining (P = 0.007). Moreover, MMP-9 results correlated with the number of obstructed meibomian ducts (P = 0.005) and a pathologic meibomian gland secretion (P = 0.001). The MMP-9 results were increased significantly in women (P < 0.001) and in patients with autoimmune disease (P = 0.005), especially Sjögren's syndrome (P = 0.001) and thyroid disease (P = 0.012).Matrix metalloproteinase 9 testing in DED is a valuable new diagnostic tool. It correlated well with other dry eye tests and identified the presence of ocular surface inflammation in 40% of confirmed dry eye patients. It may be especially helpful to identify patients with ocular surface inflammation and autoimmune disease and may facilitate the decision to institute anti-inflammatory treatment in these patients.