Characteristic cytokine profile of the aqueous humor in eyes with congenital cataract and pre-existing posterior capsule dysfunction.

Objectives: To investigate the characteristic cytokine profile of the aqueous humor in eyes with congenital cataract and pre-existing posterior capsule dysfunction (PCD). Methods: In this cross-sectional study, the enrolled eyes with congenital cataract and PCD were included in the PCD group, while those with an intact posterior capsule were included in the control group. Demographic data and biometric parameters were recorded. The levels of 17 inflammatory factors in the aqueous humor collected from the enrolled eyes were detected using Luminex xMAP technology, and intergroup differences in the collected data were analyzed. Results: The PCD group comprised 41 eyes from 31 patients with congenital cataract and PCD, whereas the control group comprised 42 eyes from 27 patients with congenital cataract and an intact posterior capsule. Lens thickness was significantly thinner in the PCD group than in the control group. However, the levels of monocyte chemoattractant protein-1 (MCP-1), transforming growth factor-ß2 (TGF-ß2), and vascular endothelial growth factor (VEGF) were significantly higher in the PCD group than in the control group. Multivariate logistic regression confirmed that lens thickness and TGF-ß2 level were independent risk factors for PCD. Conclusion: A thinner lens thickness in eyes with congenital cataract and PCD could serve as a biometric feature of these eyes. The higher levels of MCP-1, TGF-ß2, and VEGF in eyes with PCD indicated a change in their intraocular inflammatory microenvironment, which possibly led to cataract progression. Lens thickness and TGF-ß2 level are independent risk factors for PCD.

Effect of capsular bend on the rotational stability of toric intraocular lens.

PURPOSE: To assess the effect of capsular bend and axial length on the rotational stability of toric IOL. METHODS: The prospective study included patients with preexisting astigmatism that were implanted with Acrysof IQ Toric IOL. According to the pre­operative axial length, all patients were divided into 2 groups: high myopia (AL ≥ 26 mm) group, and emmetropia or low to moderate myopia group (AL <26 mm). High-speed Swept-source Optical Coherence Tomography (SS-OCT) radial scanning was performed after pupil dilation to obtain the toric IOL axial orientation and capsular bending index (CBI) at 1-day, 1-week, 1-month and 3-month intervals postoperatively. The correlation between the rotation of toric intraocular lens and the axial length or CBI was subsequently analyzed. RESULTS: 68 eyes of 57 patients were included in the research. The rotation of toric IOL within the high myopia group was greater than the control group (P = 0.001, 1month postoperative). Capsular contact with the IOL was delayed in highly myopic eyes, although the results were not statistically significant (P = 0.094, 1-month postoperatively). There was a positive correlation between the degree of rotation and axial length at the interval found between 1-week and 1-month after the operation (r = 0.333, P = 0.005). There was a significant negative correlation between the IOL rotational speed and CBI (P < 0.001). The regression equation was Y = -0.441*X + 1.712 (R2 = 0.323, P < 0.001). CONCLUSION: There was a significant negative correlation between the IOL rotation speed and the CBI, while the influence of the axial length and capsular bending mainly occurred between one week and one month after the operation.

[Preparation and identification of neutralizing antibodies to envelope protein domain III(ED III) of the four dengue virus serotypes].

Objective To prepare neutralizing monoclonal antibodies (mAbs) against envelope protein domain III of the four dengue virus serotypes (DENV-EDIII) and identify its specificity. Methods BALB/c mice were immunized with recombinant EDIII protein (rDENV-EDIII) of the four DENV serotypes. Hybridoma cells secreting DENV-EDIII antibodies were screened by indirect ELISA. The specificity of positive hybridoma cells were further tested by indirect immunofluorescence assay (IFA). The neutralizing activities of DENV-EDIII mAbs in vitro were determined by the enzyme-linked immunospot microneutralization test (ELISPOT-MNT). Results 6 mAbs specific for the EDIII of the four DENV serotypes and 11 mAbs specific for only one serotype of DENV-EDIII protein were obtained, of which one monoclonal antibody had a balanced strong neutralizing activity against the four DENV serotypes in vitro, and its 50% inhibitory concentrations (IC50) for the four DENV serotypes was 0.05 µg/mL, 1.89 µg/mL, 0.02 µg/mL, 3.91 µg/mL, respectively. Conclusion A specific monoclonal antibody against DENV-EDIII is successfully screened and obtained to neutralize the four DENV serotypes.

Posterior Capsular Outcomes of Pediatric Cataract Surgery With In-The-Bag Intraocular Lens Implantation.

Aim: To investigate the change of posterior capsular outcomes of pediatric cataract surgery with primary in-the-bag intraocular lens (IOL) implantation. Methods: We conducted a case series of pediatric cataract children who underwent cataract extraction with primary in-the-bag IOL implantation, posterior capsulorhexis or vitrectorhexis, and limited anterior vitrectomy at the Eye Hospital of Wenzhou Medical University between 2016 and 2019. Digital retro-illumination photographs of pediatric eyes were obtained at baseline and 6 months, 12 months, and the last visit postoperatively. Capsular outcomes of the posterior capsular opening area (PCOA) and lens reprolifration area at those time points were compared. Correlations between the PCOA and influential factors, such as age at surgery, axial growth, and follow-up duration, were analyzed. The study was registered at register.clinicaltrials.gov (NCT04803097). Results: Data of 23 patients (27 eyes) were used in the final analysis. During follow-up, the PCOA enlarged at a rate of 0.29-0.32 mm2/month during the first six months postoperatively and 0.05-0.08 mm2/month over the next 1-2 years. Six months postoperatively, the PCOA enlargement statistically and positively correlated with the follow-up duration and axial growth. The area of lens reprolifration was 0.46 ± 1.00 mm2 at six months postoperatively and then remained stable. Conclusion: The PCOA enlarged rapidly within the first six months after the pediatric cataract surgery with primary IOL implantation. Six months postoperatively, the enlargement of PCOA was positively correlated with follow-up duration and axial growth. Posterior capsulorhexis or capsulectomy should be performed with a diameter of 3.0 to 4.0 mm for good visual axis transparency and the protection of in-the-bag IOL.

The Relationship Between the Change of Intraocular Lens Position and Capsular Bend After Cataract Surgery.

PURPOSE: To explore the relationship between the change in intraocular lens (IOL) position and capsular bend after cataract surgery. METHODS: Patients underwent phacoemulsification and IOL implantation (Alcon Laboratories, Inc). Patients were divided into two groups based on preoperative axial length: long axial length group (axial length ⩾ 26 mm) and normal axial length group (axial length > 22 but < 26 mm). Swept-source optical coherence tomography was performed at 1 day, 1 week, 1 month, and 3 months after mydriasis to obtain postoperative aqueous depth (PAD) and capsular bend index (CBI). The relationship between CBI and PAD changes was analyzed. RESULTS: Eighty patients (80 eyes) were included in the study. PAD decreased gradually from 1 day to 1 week and increased from 1 week to 3 months. Mean CBI was moderately positively correlated with PAD changes (r = 0.586, P < .001). The IOL moved forward gradually when the CBI was less than 2.30 and the IOL gradually moved backward when the CBI was 2.30 or greater. The root mean square of the change in PAD was smaller in the long axial length group (0.08 ± 0.04 mm) than in the normal axial length group (0.09 ± 0.05 mm) during the 3 months after surgery (P = .036). CONCLUSIONS: The position of the IOL was almost stable 1 month after operation, and postoperative capsule adhesion mainly occurred within 1 month. The change in PAD was related to capsule adhesion. The postoperative position of the IOL was relatively stable and capsular bend was relatively slow for the long axial length group over 3 months. [J Refract Surg. 2021;37(5):324-330.].

Expression Profiles of Inflammatory Cytokines in the Aqueous Humor of Children after Congenital Cataract Extraction.

Purpose: To investigate the change in cytokine microenvironment of the aqueous humor (AH) after surgery in children with congenital or developmental cataracts. Methods: AH samples were obtained from 59 eyes diagnosed with a congenital or developmental cataract. Thirty-three of these eyes were aphakic following previous cataract surgery and were scheduled for secondary intraocular lens (IOL) implantation. Additionally, AH samples from 26 eyes that had not undergone surgery were taken. AH samples were then analyzed for 16 different inflammatory immune mediators using multiplex bead immunoassays and enzyme-linked immunosorbent assay (ELISA). Results: The mean interval between secondary IOL implantation and original cataract surgery was 24.85 months (range, 9-60 months). Levels of IL-6, IP-10 (CXCL10), MCP-1 (CCL2), and IL-2 were significantly elevated in the AH of eyes after surgery compared to eyes that did not undergo surgery (P < 0.001, P = 0.047, P = 0.006, P = 0.012, respectively). There was significant correlation between the levels of TGF-ß2 and intraocular pressure (IOP) in postsurgical and nonsurgical eyes (r = 0.532, P = 0.006; r = 0.57, P = 0.001). Postsurgical outcomes, such as iris adhesions, capsular fibrosis, and capsular contraction, were found not to be significantly associated with cytokine levels in the AH after surgery; however, IL-6 levels in capsular exposure eyes were significantly higher than those in cortical closure eyes (P = 0.023). Conclusions: To our knowledge, this is the first study to report significantly increased proinflammatory cytokine levels in the AH after congenital cataract extraction in children. Our study also suggests that this proinflammatory state may be maintained for a prolonged period of time. Overall, these results give us insight into the relationship between the inflammatory cytokine microenvironment of the aqueous humor and potential long-term complications following congenital cataract surgery. Translational Relevance: The inflammatory cytokine microenvironment of the aqueous humor might help explain potential long-term complications after surgery in patients with congenital or developmental cataracts.

Three-Dimensional Assessments of Intraocular Lens Stability With High-Speed Swept-Source Optical Coherence Tomography.

PURPOSE: To evaluate the effect of intraocular lens (IOL) optic overlapping on IOL stability and to determine the relationship between the capsulorhexis and IOL movement with the three-dimensional method of swept-source optical coherence tomography (SS-OCT). METHODS: This study identified patients with age-related cataracts and divided them into two groups according to their anterior capsule and IOL optic relationship: total anterior capsule overlap (360°) and partial anterior capsule overlap (< 360°). Standard SS-OCT radial scanning was performed in all eyes at 1 day, 1 week, 1 month, and 3 months after cataract surgery, respectively. The obtained photographs were used for the postoperative position measurements of capsulorhexis and IOL after three-dimensional reconstruction. RESULTS: This study included 46 eyes of 34 patients: total overlap group (n = 29) and partial overlap group (n = 17). The postoperative aqueous depth significantly decreased in the first week after surgery (P < .001). The IOL tilt was greater in the partial overlap group than that in the total overlap group (P = .014). The IOL moved significantly in the first week postoperatively (both P < .001). IOL decentration in the x-axis was greater in the partial overlap group than that in the total overlap group (P = .024). The IOL and capsulorhexis both moved sharply in the first week (both P < .05). The IOL moved consistently with the capsulorhexis in the 3 months after surgery (all P > .05). CONCLUSIONS: The total overlap group showed better IOL centrality and stability. IOL movement may be driven by capsular bag contraction and fibrosis. Thus, it was demonstrated that postoperative IOL position and IOL performance were closely linked with proper size of central continuous curvilinear capsulorhexis. [J Refract Surg. 2020;36(6):388-394.].

Longitudinal changes of the macular structure after lens removal combined with anterior vitrectomy after pediatric cataract surgery.

PURPOSE: To explore the macular thickness changes after lens removal combined with anterior vitrectomy for pediatric cataract surgery. SETTING: The Eye Hospital of Wenzhou Medical University, Zhejiang, China. DESIGN: Prospective study. METHODS: Thirty children (40 eyes) aged between 3 years and 9 years with pediatric cataracts, including 20 children (20 eyes) with unilateral cataracts and 10 children (20 eyes) with bilateral cataracts, were enrolled. Spectral-domain optical coherence tomography was used to obtain macular images. Central subfield thickness (CST) and retinal thickness in the Early Treatment Diabetic Retinopathy Study subfields (inner 1.0 to 3.0 mm annulus and outer 3.0 to 6.0 mm annulus) were recorded preoperatively as well as at 1, 3, 6, and 12 months postoperatively. RESULTS: Forty eyes of 30 children were included. Retinal thickness in every subfield significantly thickened at 3 months postoperatively (all P < .05). CST significantly thickened compared with preoperative levels (228.03 ± 18.58 vs 240.35 ± 17.41, P = .005) at 3 months postoperatively; macular thickness gradually decreased in the following months. At 6 months postoperatively, retinal thickness in inner nasal, inferior, temporal, and outer nasal subfields remained significantly thicker compared with preoperative levels (P = .048, P = .036, P = .029, and P = .017, respectively). At 12 months, the retinal thickness in all subfields reached the preoperative level. CONCLUSIONS: The influence on macular thickness lasted until 12 months after pediatric cataract surgery. The inner macular thickness increased for a longer time than the outer macular thickness postoperatively.

Clinical Characteristics and Surgical Safety in Congenital Cataract Eyes with Three Pathological Types of Posterior Capsule Abnormalities.

PURPOSE: To observe the clinical characteristics of 3 pathological types of posterior capsule abnormalities (PCAs) in congenital cataracts (CCs) and evaluate the surgical safety in these eyes. METHODS: This study involved 239 children (367 eyes) with CC who underwent cataract surgery at the Eye Hospital of Wenzhou Medical University. All surgery videos were collected for detailed reviews. Intraoperative and postoperative complications (within 3 months) were all recorded. RESULTS: The 3 pathological types of PCAs, namely, persistent fetal vasculature (PFV), posterior capsule defect (PCD), and posterior lenticonus (PLC), presented in 129 (35.1%) CC eyes, while 238 (64.9%) eyes were recorded as CC without PCA. The percentages of PFV, PCD, and PLC were 10.9%, 26.7%, and 5.4% in CC eyes (n = 367), respectively. The most common concomitant of PFV eyes was PCD (42.5%), and PFV was the most frequent (17.3%) one in PCD eyes. PLC was only associated with PFV (15%) and PCD (50%). The occurrence rates of surgical complications ranged from 0 to 5.4%, and no statistical difference was found between the eyes with and without PCA (all P > 0.05). CONCLUSIONS: PFV, PCD, and PLC play a very important role in the CCs. The effect of fetal vessels in PFV eyes might be an abnormally strong attachment on the posterior capsule, leading to PLC and PCD. Even in PCA patients, severe surgical complication can also be avoided with well-designed and skilled operation. This trial is registered with NCT03905044 at http://ClinicalTrials.gov.

Assessment of population susceptibility to upcoming seasonal influenza epidemic strain using interepidemic emerging influenza virus strains.

Seasonal influenza virus epidemics have a major impact on healthcare systems. Data on population susceptibility to emerging influenza virus strains during the interepidemic period can guide planning for resource allocation of an upcoming influenza season. This study sought to assess the population susceptibility to representative emerging influenza virus strains collected during the interepidemic period. The microneutralisation antibody titers (MN titers) of a human serum panel against representative emerging influenza strains collected during the interepidemic period before the 2018/2019 winter influenza season (H1N1-inter and H3N2-inter) were compared with those against influenza strains representative of previous epidemics (H1N1-pre and H3N2-pre). A multifaceted approach, incorporating both genetic and antigenic data, was used in selecting these representative influenza virus strains for the MN assay. A significantly higher proportion of individuals had a ⩾four-fold reduction in MN titers between H1N1-inter and H1N1-pre than that between H3N2-inter and H3N2-pre (28.5% (127/445) vs. 4.9% (22/445), P < 0.001). The geometric mean titer (GMT) of H1N1-inter was significantly lower than that of H1N1-pre (381 (95% CI 339-428) vs. 713 (95% CI 641-792), P < 0.001), while there was no significant difference in the GMT between H3N2-inter and H3N2-pre. Since A(H1N1) predominated the 2018-2019 winter influenza epidemic, our results corroborated the epidemic subtype.

The Effect of Anterior Capsule Polishing on Capsular Contraction and Lens Stability in Cataract Patients with High Myopia.

PURPOSE: To evaluate the effect of anterior capsule polishing in patients with high myopia after cataract surgery. SETTING: The Eye Hospital of Wenzhou Medical University, Zhejiang, China. DESIGN: Prospective study. METHODS: High myopic patients with a bilateral cataract who underwent phacoemulsification with 360° anterior capsular polishing in one eye and without polishing in the contralateral eye were recruited. The following parameters were recorded at 1, 3, and 6 months postoperatively, including the area and diameter of the anterior capsule opening (area and D), IOL tilt and decentration, refraction, and postoperative aqueous depth (PAD). RESULTS: Paired samples of 38 eyes of 19 patients were enrolled. The area decreased significantly in both the polished group and unpolished group, whereas the diameter reduced more in the unpolished group. The IOL tilt and decentration at 3-month and 6-month follow-up showed significant differences between two groups. In the unpolished group, the IOL decentration firstly appeared between one-month to three-month visit, while the refraction error, PAD, and IOL tilt were significantly different between the three-month and six-month visits. CONCLUSION: 360° anterior capsule polishing can effectively reduce the extent of the anterior capsule contraction and increase the stability of IOL. The study was registered at http://www.clinicaltrials.gov, and the clinical trial accession number is NCT 03142269.

Morphological and biometric features of preexisting posterior capsule defect in congenital cataract.

PURPOSE: To describe the diagnostic signs and biometric characteristics of preexisting posterior capsule defects in congenital cataract. SETTING: Eye Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China. DESIGN: Retrospective case series. METHODS: Eyes with congenital cataract were reviewed and divided into a preexisting posterior capsule defect group (study group) and a control group with intact posterior capsules. Diagnostic signs were summarized on agreement of 3 ophthalmologists after they reviewed surgical videos. The biometric parameters, including central corneal thickness, anterior chamber depth, lens thickness, vitreous chamber depth, axial length, mean keratometry, corneal astigmatism, corneal diameter, and intraocular pressure, were compared between groups. The predictors of preexisting posterior capsule defect were determined using univariate and multivariate analyses. RESULTS: The study comprised 81 eyes (53 patients). The diagnostic signs of preexisting posterior capsule defect included posterior capsule plaque with a well-demarcated margin, gray granules and vacuoles, and heterogeneous nuclear opacity (more dense in the posterior). The lens was significantly thinner in the study group than in the control group (P < .001). The corneal diameter was significantly smaller in the study group (P = .015 horizontal and P = .005 vertical). Multivariate analysis identified lens thickness as an independent predictor (odds ratio, 0.731; 95% confidence interval, 0.621-0.860; P < .001). CONCLUSIONS: For preexisting posterior capsule defect, lens thickness was a valuable predictor and the presence of characteristic morphology was an important indicator. The cornea configuration suggests the need for further study of the role of genetic mutation.

Functional properties of DENV EDIII­reactive antibodies in human DENV­1­infected sera and rabbit antiserum to EDIII.

The envelope domain III (EDIII) of the dengue virus (DENV) has been confirmed to be involved in receptor binding. It is the target of specific neutralizing antibodies, and is considered to be a promising subunit dengue vaccine candidate. However, several recent studies have shown that anti­EDIII antibodies contribute little to the neutralizing or enhancing ability of human DENV­infected serum. The present study involved an analysis of the neutralization and antibody­dependent enhancement (ADE) activities of EDIII­reactive antibodies in human convalescent sera from patients with primary DENV­1 infection and rabbit antiserum immunized with recombinant DENV­1 EDIII protein. The results indicated that serum neutralization was not associated with titres of EDIII­binding antibodies in the human DENV­1­infected sera. The depletion of anti­EDIII antibodies from these serum samples revealed that the anti­EDIII antibodies of the patients contributed little to neutralization and ADE. However, the EDIII­reactive antibodies from the rabbit antiserum exhibited protective abilities of neutralization at a high dilution (~1:50,000) and ADE at a low dilution (~1:5,000) for the homotypic DENV infection. Notably, the rabbit antiserum displayed ADE activity only at a dilution of 1:40 for the heterotypic virus infection, which suggests that EDIII­reactive antibodies may be safe in secondary infection with heterotypic viruses. These results suggest that DENV EDIII is not the predominant antigen of the DENV infection process; however, purified or recombinant DENV EDIII may be used as a subunit vaccine to provoke an effective and safe antibody response.

An Optical Section-Assisted In Vivo Rabbit Model for Capsular Bend and Posterior Capsule Opacification Investigation.

PURPOSE: To establish an optical section-assisted in vivo rabbit model for capsular bend and posterior capsule opacification (PCO) investigation. METHODS: A total of 10 rabbits underwent phacoemulsification surgery and intraocular lens (IOL) implantation. On the basis of the relationship between the anterior capsule and IOL, the rabbits were divided into complete overlap and incomplete overlap groups, in which six and four rabbits were included, respectively. The capsular bend optical sections were assessed using ultra-long scan depth optical coherence tomography (UL-OCT), and posterior capsule opacification was evaluated with slit lamp on postoperative day 3, 7, 14, and 28. In addition, histopathological section was used to verify the accuracy of capsular bend type captured by OCT in three rabbits. RESULTS: Based on the special animal model, six capsular bend types were observed, namely, anterior (A), middle (M), posterior (P), detachment (D), funnel (Fun) and furcate adhesion (Fur). On day 3, capsular bend began to form. On 14 days, the capsular bends were comprised of A, M and D types, which were almost maintained until day 28. Histopathological section findings were consistent with optical sectioning results. In the incomplete and complete groups, the earliest PCO within the optical zone were on day 7 and 28, respectively. The incomplete group exhibited higher incidence and faster PCO on day 7 (p = 0.038) and 14 (p = 0.002). CONCLUSIONS: This animal model not only mimics capsular bend evolution and PCO processes but also produces OCT optical section images equivalent to and more repeatable than histopathology, thereby providing a promising method for the further investigations of PCO.

The Repeatability Assessment of Three-Dimensional Capsule-Intraocular Lens Complex Measurements by Means of High-Speed Swept-Source Optical Coherence Tomography.

PURPOSE: To rebuild the three-dimensional (3-D) model of the anterior segment by high-speed swept-source optical coherence tomography (SSOCT) and evaluate the repeatability of measurement for the parameters of capsule-intraocular lens (C-IOL) complex. METHODS: Twenty-two pseudophakic eyes from 22 patients were enrolled. Three continuous SSOCT measurements were performed in all eyes and the tomograms obtained were used for 3-D reconstruction. The output data were used to evaluate the measurement repeatability. The parameters included postoperative aqueous depth (PAD), the area and diameter of the anterior capsule opening (Area and D), IOL tilt (IOL-T), horizontal, vertical, and space decentration of the IOL, anterior capsule opening, and IOL-anterior capsule opening. RESULTS: PAD, IOL-T, Area, D, and all decentration measurements showed high repeatability. Repeated measure analysis showed there was no statistically significant difference among the three continuous measurements (all P > .05). Pearson correlation analysis showed high correlation between each pair of them (all r >0.90, P<0.001). ICCs were all more than 0.9 for all parameters. The 95% LoAs of all parameters were narrow for comparison of three measurements, which showed high repeatability for three measurements. CONCLUSION: SSOCT is available to be a new method for the 3-D measurement of C-IOL complex after cataract surgery. This method presented high repeatability in measuring the parameters of the C-IOL complex.

A comparison of Luminex xTAG® Gastrointestinal Pathogen Panel (xTAG GPP) and routine tests for the detection of enteropathogens circulating in Southern China.

To investigate whether Luminex xTAG® Gastrointestinal Pathogen Panel (xTAG GPP) is applicable for the diagnosis of diarrhea and surveillance of enteropathogens circulating in Southern China, 290 stool samples were tested for 15 kinds of enteropathogens using xTAG GPP and compared to the results from the routine tests, including culture; immunochromatography; real-time PCR; microscopy; and a third method, gene sequencing. One hundred fifty-nine samples were positive, yielding a total of 181 enteropathogens (69 bacteria and 112 viruses), with rotavirus being most prevalent (39.0%, 62/159). The overall sensitivity and specificity of xTAG GPP were 96.3% (93.3-98.2%) and 99.8% (99.6-99.9%), respectively, with a combination of the methods as the gold standard. The coinfection rates detected by the routine tests and xTAG GPP were 10.0% (25 double and 4 triple infections) and 12.1% (29 double, 4 triple and 2 quadruple infections), respectively. xTAG GPP is a powerful tool for the identification of multiple enteropathogens.

Anterior chamber depth measurements using Scheimpflug imaging and optical coherence tomography: repeatability, reproducibility, and agreement.

PURPOSE: To determine the repeatability, reproducibility, and agreement of anterior chamber depth (ACD) measurements obtained with 3 Scheimpflug cameras and an anterior segment optical coherence tomography (AS-OCT) device. SETTING: Eye Hospital of Wenzhou Medical University, Zhejiang, China. DESIGN: Observational cross-sectional study. METHODS: Two observers took 3 consecutive measurements in healthy right eyes using each device to assess intraoperator repeatability. The mean values obtained at different sessions by the first operator were used to determine the intersession reproducibility. Three consecutive measurements obtained by the first operator at the first session were averaged and used to assess agreement. RESULTS: The ACD measurements obtained by 2 observers in 71 eyes were highly repeatable using the 4 devices, with a test-retest repeatability of 0.04 to 0.07 mm for intraoperator repeatability. The interoperator and intersession reproducibility of ACD measurements were high, and the test-retest repeatability ranges of interoperator and intersession reproducibility were 0.06 to 0.07 mm and 0.05 to 0.08 mm, respectively. The ACD measurements of the 4 systems were sorted from the thickest to the thinnest (Galilei G2 > Visante > Sirius > Pentacam). The differences in the measurements were statistically significant except between the ACD measurements obtained by the Sirius device and the Visante device. However, good agreement with narrow 95% limits of agreement was found between these devices. CONCLUSIONS: The 4 devices provided high intraoperator repeatability and interoperator and intersession reproducibility for ACD measurements. Good agreement in ACD measurements was found between the devices in healthy eyes. FINANCIAL DISCLOSURE: No author has a financial or proprietary interest in any material or method mentioned.

Development of a double antibody sandwich ELISA for West Nile virus detection using monoclonal antibodies against non-structural protein 1.

The early diagnosis of West Nile virus (WNV) infection is important for successful clinical management and epidemiological control. The non-structural protein 1 (NS1) of flavivirus, a highly conserved and secreted glycoprotein, is abundant in the serum of flavivirus-infected patients and represents a useful early diagnostic marker. We developed a WNV-specific NS1 antigen-capture ELISA using two mouse monoclonal antibodies (MAbs) that recognised distinct epitopes of the NS1 protein of WNV as capture and detection antibodies. The antigen-capture ELISA displayed exclusive specificity to WNV without cross-reaction with other related members of the flavivirus family, including the dengue virus, yellow fever virus, Japanese encephalitis virus, and tick-borne encephalitis virus. Additionally, the specificity was presented as no false positive in normal (0/1003) and DENV-infected (0/107) human serum specimens. The detection limit of the antigen-capture ELISA was as low as 15 pg/ml of recombinant WNV NS1 protein (rWNV-NS1) and 6.1 plaque-forming units (PFU)/0.1 ml of WNV-infected culture supernatant. In mice infected with WNV, the NS1 protein was readily detected in serum as early as one day after WNV infection, prior to the development of clinical signs of the disease. The sensitivity of the NS1 capture ELISA (93.7%) was significantly higher (79.4%) than that of real-time reverse transcription polymerase chain reaction in 63 serum samples from WNV-infected mice (p = 0.035). This newly developed NS1 antigen-capture ELISA with high sensitivity and specificity could be used as an efficient method for the early diagnosis of WNV infection in animals or humans.

The absence of exanthema is related with death and illness severity in acute enterovirus infection.

OBJECTIVE: To clarify whether exanthema is related to illness severity in acute enterovirus infection in children. METHODS: The data of pediatric inpatients at Zhujiang Hospital during 2009-2012 with an acute enterovirus infection were reviewed retrospectively. Enterovirus infection was determined by real-time reverse transcription PCR. Clinical data were summarized and compared between cases with and without exanthema. RESULTS: A total of 780 pediatric inpatients with an acute enterovirus infection were included in this study, of whom 83 (10.6%) presented no exanthema. The percentage of deaths in the group of patients without exanthema was significantly higher than that in the group with exanthema (7.2% vs. 1.1%; p = 0.002). Central nervous system involvement (41.0% vs. 30.0%; p = 0.041), severe central nervous system (CNS) involvement (21.7% vs. 11.0%; p = 0.005), severe CNS involvement with cardiopulmonary failure (9.6% vs. 2.3%; p = 0.002), an altered level of consciousness (15.7% vs. 7.6%; p = 0.013), and convulsions (14.4% vs. 6.3%; p = 0.007) occurred significantly more frequently in the group without exanthema. CONCLUSIONS: A considerable proportion of children with an acute enterovirus infection in Guangdong Province, China during 2009-2012 presented no exanthema, and the absence of exanthema was found to be related to death and illness severity for these acute enterovirus infections. Clinicians in China should consider enterovirus as the possible pathogen when treating children with an acute pathogen infection without exanthema.

Comparison of MALDI-TOF MS, gene sequencing and the Vitek 2 for identification of seventy-three clinical isolates of enteropathogens.

OBJECTIVE: This study was performed to evaluate the analytical and practical performance of the matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) compared to the sequencing method and the Vitek 2 system for identification of enteropathogens in the clinical microbiology laboratory. METHODS: Ten type strains and 73 clinical isolates of enteropathogens representing eight genera were analyzed by MALDI-TOF MS. All isolates were also characterized by gene sequencing allowing interpretation of the results from MALDI-TOF MS. In addition, MALDI-TOF MS was compared with the Vitek 2 system for the identification of ten isolates of Aeromonas and six of Salmonella. RESULTS: As previously known, identification between Shigella and Escherichia coli is not possible to distinguish. MALDI-TOF MS produced the correct identifications for all other type strains and clinical isolates to the genus level. Fifteen Campylobacter jejuni, six Campylobacter coli, three Plesiomonas shigelloides, three Yersinia enterocolitica, two Clostridium difficile, one Vibrio parahaemolyticus, one Vibrio fluvialis, and one Vibrio cholera were all correctly identified to the species level. Genus and species identifications of ten Aeromonas and six Salmonella isolates by MALDI-TOF MS were consistent with those by the Vitek 2, but with much less cost and about ten times faster. CONCLUSIONS: This study demonstrates that MALDI-TOF MS is a powerful tool for fast, accurate and low-cost identification of enteropathogens in the clinical microbiology laboratory.