@article {1056, title = {Deep sequencing analysis of clinical samples from patients with acute infectious conjunctivitis during the COVID-19 delta surge in Madurai, India.}, journal = {J Clin Virol}, volume = {157}, year = {2022}, month = {2022 Oct 05}, pages = {105318}, abstract = {

BACKGROUND: Seasonal outbreaks of infectious conjunctivitis remain a public health issue. Determination of outbreak etiologies in the context of a worldwide pandemic may provide useful information to guide public health strategies. The aim of this study was to identify pathogens associated with outpatient infectious conjunctivitis during the COVID-19 Delta surge.

METHODS: This prospective study was conducted from April 2021 to September 2021. All outpatients presenting to the Aravind Eye Center (Madurai, India) with signs and symptoms consistent with acute infectious conjunctivitis were eligible. Three swabs were obtained from each participant: one from each conjunctiva and one from the anterior nares. Samples were processed for metagenomic RNA deep sequencing (RNA-seq).

RESULTS: Samples from 106 study participants were sequenced. The most common presenting symptoms were tearing (86\%) and itching (71\%). Preauricular lymphadenopathy was present in 38\% of participants. 20\% of participants had close contacts with similar symptoms. Systemic symptoms such as coughing, runny nose, vomiting or diarrhea were uncommonly reported. 60\% of all participants used some medicated eye drops upon enrollment. 75\% of study participants demonstrated infection with human adenovirus D (HAdV-D). 11\% of conjunctivitis was associated with SARS-CoV-2. 15\% had no definitive pathogen detected. 8\% of all participants had codetection of more than one pathogen on RNA-seq.

CONCLUSIONS: During the COVID-19 Delta surge in India, HAdV-D was the most common pathogen associated with infectious conjunctivitis. SARS-CoV-2 was the second most common associated pathogen. Seasonal surveillance may be necessary for the determination of emerging and reemerging pathogens responsible for infectious conjunctivitis.

}, issn = {1873-5967}, doi = {10.1016/j.jcv.2022.105318}, author = {Lalitha, Prajna and Prajna, N Venkatesh and Gunasekaran, Rameshkumar and Teja, Gonugunta Vishnu and Sharma, Sankalp S and Hinterwirth, Armin and Ruder, Kevin and Zhong, Lina and Chen, Cindi and Deiner, Michael and Huang, ChunHong and Pinsky, Benjamin A and Lietman, Thomas M and Doan, Thuy and Seitzman, Gerami D} } @article {966, title = {Interepidemic Respiratory Syncytial Virus during the COVID-19 Pandemic.}, journal = {Microbiol Spectr}, year = {2022}, month = {2022 Apr 25}, pages = {e0094722}, issn = {2165-0497}, doi = {10.1128/spectrum.00947-22}, author = {Huang, ChunHong and Sahoo, Malaya K and Verghese, Michelle and Sibai, Mamdouh and Solis, Daniel and Mfuh, Kenji O and Kurzer, Jason and Hogan, Catherine A and Doan, Thuy A and Pinsky, Benjamin A} } @article {1046, title = {Outpatient human coronavirus associated conjunctivitis in India.}, journal = {J Clin Virol}, volume = {157}, year = {2022}, month = {2022 Sep 27}, pages = {105300}, abstract = {

BACKGROUND: Viral conjunctivitis (pink eye) can be highly contagious and is of public health importance. There remains significant debate whether SARS-CoV-2 can present as a primary conjunctivitis. The aim of this study was to identify pathogens associated with outpatient infectious conjunctivitis during the COVID-19 Delta surge.

METHODS: This prospective study was conducted in the spring and summer months of 2021. 106 patients with acute conjunctivitis who presented to the Aravind Eye Center in Madurai, India were included. One anterior nasal swab and one conjunctival swab of each eye were obtained for each enrolled patient. Samples were subsequently processed for unbiased metagenomic RNA deep sequencing (RNA-seq). Outcomes included clinical findings and codetection of other pathogens with SARS-CoV-2 in patients with conjunctivitis.

RESULTS: Among the 13 patients identified with human coronavirus RNA fragments in their swabs, 6 patients had SARS-CoV-2 infection, 5 patients had coinfections of SARS-CoV-2 and human adenovirus (HAdV), 1 patient had a coinfection with human coronavirus OC43 and HAdV, and 1 patient had a coinfection of Vittaforma corneae and SARS-CoV-2. 30\% had bilateral disease and symptoms on presentation. Petechial hemorrhage was noted in 33\% of patients with SARS-CoV-2 infection. No patients with SARS-CoV-2 or SARS-CoV-2 and HAdV infections had subepithelial infiltrates on presentation. All patients denied systemic symptoms.

CONCLUSIONS: Among the patients presented with conjunctivitis associated with human coronavirus infection, over 50\% of the patients had co-infections with other circulating pathogens, suggesting the public-health importance of broad pathogen testing and surveillance in the outpatient conjunctivitis population.

}, issn = {1873-5967}, doi = {10.1016/j.jcv.2022.105300}, author = {Prajna, N Venkatesh and Lalitha, Prajna and Teja, Gonugunta Vishnu and Gunasekaran, Rameshkumar and Sharma, Sankalp S and Hinterwirth, Armin and Ruder, Kevin and Zhong, Lina and Chen, Cindi and Deiner, Michael and Huang, ChunHong and Pinsky, Benjamin A and Lietman, Thomas M and Seitzman, Gerami D and Doan, Thuy} } @article {946, title = {Vaccine-Associated Measles Encephalitis in Immunocompromised Child, California, USA.}, journal = {Emerg Infect Dis}, volume = {28}, year = {2022}, month = {2022 Apr}, pages = {906-908}, abstract = {

We report a fatal case of vaccine-associated measles encephalitis in an immunocompromised child in California, USA. The infection was confirmed by whole-genome RNA sequencing of measles virus from brain tissue. We observed biased matrix-gene hypermutation consistent with persistent measles virus central nervous system infection.

}, issn = {1080-6059}, doi = {10.3201/eid2804.212357}, author = {Costales, Cristina and Sahoo, Malaya K and Huang, ChunHong and Guimaraes, Carolina V and Born, Donald and Kushner, Lauren and Gans, Hayley A and Doan, Thuy A and Pinsky, Benjamin A} } @article {731, title = {Comprehensive pathogen detection for ocular infections.}, journal = {J Clin Virol}, volume = {136}, year = {2021}, month = {2021 Feb 11}, pages = {104759}, abstract = {

BACKGROUND: Molecular diagnostics such as pathogen-directed PCRs have transformed testing for ocular infections since the late 1990s. Although these assays remain important diagnostic tools for samples with low biomass, the lack of diagnostic range motivates alternative molecular approaches for ocular infections. The aim of this study was to determine the performance of a high-throughput RNA sequencing approach, RNA-seq, to detect infectious agents in ocular samples from patients with presumed ocular infections.

METHODS: We compared the performance of RNA-seq to pathogen-directed PCRs using remnant nucleic acids from 41 aqueous or vitreous samples of patients with presumed ocular infections. Pathogen-directed PCRs were performed at the CLIA-certified Stanford Clinical Virology Laboratory. RNA-seq was performed in a masked manner at the Proctor Foundation at the University of California San Francisco. Percent positive and negative agreement between the two testing approaches were calculated. Discordant results were subjected to orthogonal testing.

RESULTS: The positive percent agreement between RNA-seq and pathogen-directed PCRs was 100\% (95\% confidence interval (CI): 78.5\%-100\%). The negative percent agreement was 92.6\% (95\% CI: 76.6\%-97.9\%). RNA-seq identified pathogens not on the differential diagnosis for 9.7\% (4/41) of the samples. Two pathogens solely identified with RNA-seq were confirmed with orthogonal testing.

CONCLUSIONS: RNA-seq can accurately identify common and rare pathogens in aqueous and vitreous samples of patients with presumed ocular infections. Such an unbiased approach to testing has the potential to improve diagnostics although practical clinical utility warrants additional studies.

}, issn = {1873-5967}, doi = {10.1016/j.jcv.2021.104759}, author = {Doan, Thuy and Sahoo, Malaya K and Ruder, Kevin and Huang, ChunHong and Zhong, Lina and Chen, Cindi and Hinterwirth, Armin and Lin, Charles and Gonzales, John A and Pinsky, Benjamin A and Acharya, Nisha R} } @article {51, title = {Metagenomic Next-Generation Sequencing for Identification and Quantitation of Transplant-Related DNA Viruses.}, journal = {J Clin Microbiol}, volume = {57}, year = {2019}, month = {2019 12}, abstract = {

Infections with DNA viruses are frequent causes of morbidity and mortality in transplant recipients. This study describes the analytical and clinical performance characteristics of the Arc Bio Galileo Pathogen Solution, an all-inclusive metagenomic next-generation sequencing (mNGS) reagent and bioinformatics pipeline that allows the simultaneous quantitation of 10 transplant-related double-stranded DNA (dsDNA) viruses (adenovirus [ADV], BK virus [BKV], cytomegalovirus [CMV], Epstein-Barr virus [EBV], human herpesvirus 6A [HHV-6A], HHV-6B, herpes simplex virus 1 [HSV-1], HSV-2, JC virus [JCV], and varicella-zoster virus [VZV]). The mNGS 95\% limit of detection ranged from 14 copies/ml (HHV-6) to 191 copies/ml (BKV), and the lower limit of quantitation ranged from 442 international units (IU)/ml (EBV) to 661 copies/ml (VZV). An evaluation of 50 residual plasma samples with at least one DNA virus detected in prior clinical testing showed a total percent agreement of mNGS and quantitative PCR (qPCR) of 89.2\% (306/343), with a κ statistic of 0.725. The positive percent agreement was 84.9\% (73/86), and the negative percent agreement was 90.7\% (233/257). Furthermore, mNGS detected seven subsequently confirmed coinfections that were not initially requested by qPCR. Passing-Bablok regression revealed a regression line of = 0.953 + 0.075 (95\% confidence interval [CI] of the slope, 0.883 to 1.011; intercept, -0.100 to 0.299), and Bland-Altman analysis (mNGS - qPCR) showed a slight positive bias (0.28 log concentration; 95\% limits of agreement, -0.62 to 1.18). In conclusion, the mNGS-based Galileo pipeline demonstrates analytical and clinical performance comparable to that of qPCR for transplant-related DNA viruses.

}, keywords = {Computational Biology, DNA Virus Infections, DNA Viruses, High-Throughput Nucleotide Sequencing, Humans, Metagenomics, Molecular Diagnostic Techniques, Sensitivity and Specificity, Transplantation}, issn = {1098-660X}, doi = {10.1128/JCM.01113-19}, author = {Carpenter, Meredith L and Tan, Susanna K and Watson, Thomas and Bacher, Rowena and Nagesh, Vaishnavi and Watts, Alain and Bentley, Gordon and Weber, Jenna and Huang, ChunHong and Sahoo, Malaya K and Hinterwirth, Armin and Doan, Thuy and Carter, Theodore and Dong, Queeny and Gourguechon, St{\'e}phane and Harness, Eric and Kermes, Sean and Radhakrishnan, Srihari and Wang, Gongbo and Quiroz-Z{\'a}rate, Alejandro and Ching, Jesus and Pinsky, Benjamin A} }