ABSTRACT
@#Abstract: Objective To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.
ABSTRACT
ObjectiveTo observe the effect of Chonglian oral liquid on inflammatory and immune markers as well as the clinical outcomes of patients with mild-to-moderate corona virus disease 2019（COVID-19） and comprehensively evaluate its efficacy and safety. MethodA clinical randomized controlled trial （RCT） was conducted， involving 120 confirmed cases of mild-to-moderate COVID-19. The patients were randomly divided into two groups， with 55 cases in the observation group and 56 cases in the control group. According to the updated diagnosis and treatment protocol， the control group received standard western medical treatment， while the observation group received Chonglian oral liquid in addition to standard western medical treatment. Both groups were treated continuously for 10 days. The traditional Chinese medicine （TCM） syndrome scores， syndrome efficacy， fever abatement time， nucleic acid negative conversion time， inflammatory and immune markers， improvement in imaging findings， clinical outcomes， and occurrence of adverse events were compared between the two groups. ResultBoth groups showed a significant decrease in TCM syndrome scores after treatment （P<0.01）. Compared with the control group after treatment， the observation group exhibited a more significant improvement in cough， dry throat， sore throat， fatigue， and muscle pain （P<0.05）. The total effective rate in the observation group was 100% （55/55）， significantly higher than 98.21% （55/56） in the control group （Z=3.707， P<0.01）. The observation group also showed a significantly shorter duration of fever abatement and nucleic acid negative conversion compared with the control group after treatment （P<0.05）. Both groups had a significant increase in lymphocyte count （LYM）， lymphocyte percentage （LYM%）， mature T lymphocytes （CD3+）， and helper/inducer T lymphocytes （CD4+） after treatment （P<0.01）. Compared with the control group after treatment， the observation group showed greater improvement in these markers （P<0.05）. In terms of inflammatory markers， both groups had a significant decrease compared with those before treatment （P<0.01）. The observation group exhibited lower levels of high-sensitivity C-reactive protein （hs-CRP）， interleukin-6 （IL-6）， and procalcitonin （PCT） than the control group after treatment （P<0.05）. There was no statistically significant difference in imaging efficacy evaluation and clinical outcomes between the two groups. No adverse events were reported in either group during the treatment period. ConclusionChonglian oral liquid combined with standard western medical treatment significantly improves clinical symptoms， shortens fever abatement and nucleic acid negative conversion time， regulates immune function， and inhibits inflammatory responses in patients with mild-to-moderate COVID-19， leading to improved clinical efficacy.
ABSTRACT
La enfermedad del corona virus 2 del síndrome respiratorio agudo severo (virus SARS-CoV-2) apareció por primera vez en diciembre de 2019 en Wuhan, China, y desde entonces se ha extendido rápidamente por todo el mundo. Desde entonces, el brote de esta grave enfermedad viral se ha convertido en una amenaza global para la humanidad. El diagnóstico precoz y el aislamiento son las medidas más importantes necesarias para prevenir su propagación. La evidencia anecdótica reciente ha sugerido manifestaciones orales con o sin deterioro olfativo y gustativo en asociación con la enfermedad por coronavirus (COVID-19). La enzima convertidora de angiotensina-2 (ECA-2) se expresa en la mucosa oral en grandes cantidades y, por tanto, puede contribuir a las primeras manifestaciones de esta enfermedad viral mortal. Las manifestaciones bucales de la enfermedad por coronavirus pueden presentarse en forma de lesiones ulcerativas irregulares en relación con diferentes partes de la cavidad oral y, en particular, en relación con la mucosa adherida en la región del paladar duro, así como inflamación y posterior atrofia de las diversas papilas de la lengua. La disfunción olfativa y gustativa asociada también puede conducir a una pérdida parcial y / o incluso completa de la capacidad para oler y saborear en las primeras etapas del inicio de la enfermedad. La evidencia también ha sugerido la presencia de ácido nucleico del SARS-CoV-2 en la saliva humana, lo que la convierte en portadora de la enfermedad viral infecciosa y ayuda en su diagnóstico. Hemos buscado sistemáticamente la base de datos médica para el mismo y hemos revisado toda la literatura disponible hasta el 29 de junio de 2020
Severe acute respiratory syndrome corona virus 2 (SARS-CoV-2 virus) disease had first appeared in December 2019 in Wuhan, China and has been spreading quickly throughout the world since then. Since then, the outbreak of this severe viral disease has become a global threat to humanity. An early diagnosis and isolation are the most significant measures required to prevent its spread. Recent anecdotal evidence has suggested oral manifestations with or, without olfactory and gustatory impairment in association with corona virus disease (COVID-19). Angiotensin converting enzyme-2 (ACE-2) is expressed in oral mucosa in large amounts and can, thus, contribute in the early manifestations of this deadly viral disease. The oral manifestations of corona virus disease can occur in the form of irregular ulcerative lesions in relation to different parts of the oral cavity and particularly, in relation to the attached mucosa in the hard palate region as well as inflammation and subsequent, atrophy of the various tongue papilla. The associated olfactory and gustatory dysfunction can, also, lead to partial and/or, even a complete loss of the ability to smell and taste in the early stages of the disease onset. Evidence has, also, suggested the presence of SARS-CoV-2 nucleic acid in human saliva making it the carrier of the infectious viral disease as well as aiding in its diagnosis. We have systemically searched medical database for the same and have reviewed all the literature available up to 29th of June 2020.