ABSTRACT
The rapid emergence of coronavirus disease 2019 (COVID-19) as a global pandemic affecting millions of individuals globally has necessitated sensitive and high-throughput approaches for the diagnosis, surveillance, and determining the genetic epidemiology of SARS-CoV-2. In the present study, we used the COVIDSeq protocol, which involves multiplex-PCR, barcoding, and sequencing of samples for high-throughput detection and deciphering the genetic epidemiology of SARS-CoV-2. We used the approach on 752 clinical samples in duplicates, amounting to a total of 1536 samples which could be sequenced on a single S4 sequencing flow cell on NovaSeq 6000. Our analysis suggests a high concordance between technical duplicates and a high concordance of detection of SARS-CoV-2 between the COVIDSeq as well as RT-PCR approaches. An in-depth analysis revealed a total of six samples in which COVIDSeq detected SARS-CoV-2 in high confidence which were negative in RT-PCR. Additionally, the assay could detect SARS-CoV-2 in 21 samples and 16 samples which were classified inconclusive and pan-sarbeco positive respectively suggesting that COVIDSeq could be used as a confirmatory test. The sequencing approach also enabled insights into the evolution and genetic epidemiology of the SARS-CoV-2 samples. The samples were classified into a total of 3 clades. This study reports two lineages B.1.112 and B.1.99 for the first time in India. This study also revealed 1,143 unique single nucleotide variants and added a total of 73 novel variants identified for the first time. To the best of our knowledge, this is the first report of the COVIDSeq approach for detection and genetic epidemiology of SARS-CoV-2. Our analysis suggests that COVIDSeq could be a potential high sensitivity assay for the detection of SARS-CoV-2, with an additional advantage of enabling the genetic epidemiology of SARS-CoV-2.
Subject(s)
COVID-19/epidemiology , COVID-19/virology , High-Throughput Nucleotide Sequencing/methods , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , COVID-19/genetics , Genome, Viral/genetics , Humans , India/epidemiology , Molecular Epidemiology/methods , Multiplex Polymerase Chain Reaction/methods , Pandemics , Phylogeny , RNA, Viral/genetics , RNA, Viral/isolation & purification , Sensitivity and SpecificityABSTRACT
BACKGROUND: In the treatment of phrynoderma, various nutrients have been tried in different dosages and routes with variable therapeutic outcomes. AIMS: The present study was undertaken to compare the efficacy of various nutrients in the treatment of phrynoderma. SETTINGS AND DESIGN: An open label randomized controlled study was conducted in the out-patient department of Dermatology in a tertiary care hospital. MATERIAL AND METHODS: The patients were divided into group of five and each patient received one of the five regimens [10 injections of Vitamin A 1 lakh units, intramuscularly (IM) on alternate day, 10 injections of Vitamin B complex, 2cc IM on alternate day, 2 table spoon of safflower oil, two times daily with meals, Tab Vitamin E 400mg once daily, and only topical keratolytic (salicylic acid 3% ointment) two times daily] respectively. The first four regimens also received topical keratolytic. The primary outcome measured was therapeutic response at the end of regimen. The response was graded based on the percentage of flattening and decrease in number of lesions. Less than 25% improvement was graded as poor, 26-50% improvement as moderate, 51-75% improvement as good, and more than 75% improvement as excellent response. In the statistical analysis, comparison was done using Chi-square and Fisher's exact test. RESULTS: A total of 125 patients were included in the study with 25 patients each in five regimen groups. There were 79 (63.2%) males and 46 (36.8%) females with a ratio of 1.72:1. The age of the patients ranged from 3 to 26 years with mean of 10.1±4.3 years. Out of 125 patients, 105 completed the study. In regimen 1 and 2, significant number of patients showed good to excellent response whereas significant number of patients in remaining regimen showed moderate to poor response with a p value of <0.05. The therapeutic response to Vitamin A and Vitamin B complex therapy was significantly better than other regimens. CONCLUSION: Intramuscular injections of Vitamin A or Vitamin B are effective in the treatment of phrynoderma. Addition of topical keratolytics helps in faster clearance of keratotic lesions.
