Objective: Most cases of coronavirus disease 2019 (COVID-19) are identified as moderate, which is defined as having a fever or dry cough and lung imaging with ground-glass opacities. The risk factors and predictors of prognosis in such cohorts remain uncertain. Methods: All adults with COVID-19 of moderate severity diagnosed using quantitative RT-PCR and hospitalized at the Central Hospital of Wuhan, China, from 1 January to 20 March 2020 were enrolled in this retrospective study. The main outcomes were progression from moderate to severe or critical condition or death. Results: Among the 456 enrolled patients with moderate COVID-19, 251/456 (55.0%) had poor prognosis. Multivariate logistic regression analysis identified higher neutrophil count: lymphocyte count ratio (NLR) on admission (OR 1.032, 95% CI 1.042e1.230, p 0.004) and higher C-reactive protein (CRP) on admission (OR 3.017, 95% CI 1.941e4.690, p < 0.001) were associated with increased OR of poor prognosis. The area under the receiver operating characteristic curve (AUC) for NLR and CRP in predicting progression to critical condition was 0.77 (95% CI 0.694e0.846, p < 0.001) and 0.84 (95% CI 0.780e0.905, p < 0.001), with a cutoff value of 2.79 and 25.95 mg/L, respectively. The AUC of NLR and CRP in predicting death was 0.81 (95% CI 0.732e0.878, p < 0.001) and 0.89 (95% CI 0.825e0.946, p < 0.001), with a cutoff value of 3.19 and 33.4 mg/L, respectively. Conclusions: Higher levels of NLR and CRP at admission were associated with poor prognosis of individuals with moderate COVID-19. NLR and CRP were good predictors of progression to critical condition and death.

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Hand, foot, and mouth disease (HFMD) is an infectious disease caused by human enterovirus 71 (EV71), coxsackievirus A16 (CVA16) and other enteroviruses. It is of interest that other enteroviruses associated with HFMD in Jinan have been rarely reported. The aim of the present study is to detect and characterize the circulating serotypes of non-EV71 and non-CVA16 enteroviruses associated with HFMD in Jinan city, Shandong province, China. A total of 400 specimens were collected from clinically diagnosed HFMD cases in Jinan from January 2009 to June 2013. All specimens were infected with non-EV71 and non-CVA16 enteroviruses previously confirmed by RT-PCR or real-time PCR according to the protocols at that time. The GeXP-based multiplex RT-PCR assay (GeXP assay) was performed to investigate the pathogen spectrum of 15 enteroviruses (coxsackieviruses A4, A5, A6, A9, A10, A16; coxsackieviruses B1, B3, B5; Echoviruses 6, 7, 11, 13, 19 and EV71) infections associated with HMFD. For GeXP assay negative samples, reverse transcription nested PCR (nested RT-PCR) based on the 5’ -untranslated region (5’- UTR) sequence and phylogenetic analysis were conducted to further explore the etiology of multiple enteroviruses. The results showed that a total of twenty serotypes of enteroviruses (including EV71 and CVA16) were identified by GeXP assay and nested RT-PCR. The most circulating twelve serotypes of enteroviruses with HFMD in Jinan from 2009 to June 2013 were EV71, CVA16, CVA10, CVA6, CVA12, CVA2, Echo3, CVA4, CVA9, CVB1, CVB3 and Echo6. CVA10 and CVA6 were the most prevalent pathogens other than EV71 and CVA16 in Jinan and their most prevalent seasons were spring and summer, and a slight increase was observed in autumn and early winter. It should be noted that mixed-infections were identified by GeXP assay and the phylogenetic tree clearly discriminated the multiple pathogens associated with HFMD. Our results thus demonstrate that there was a clear lack of a reliable testing method for EV71 and CVA16 and multiple non-EV71 and non-CVA16 enteroviruses associated with HFMD were present in Jinan. The GeXP assay combined with nested RT-PCR based on 5’-UTR region could meet the need for the national surveillance of multiple enteroviruses or the investigation of epidemic outbreaks triggered by enteroviruses in the future.

The etiological agent of severe fever with thrombocytopenia syndrome (SFTS) is a bunyavirus that was first identified in China in 2009. We have developed and validated a one-step, single-tube, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of SFTS bunyavirus (SFTSV). This assay demonstrated high specificity and sensitivity, with a detection limit of 10(1) TCID(50) ml(-1). When combined with the fluorescent detection reagent (FDR) method, results could be determined by observing a color change within 30 min. As an accurate, rapid, simple and low-cost diagnostic method, this RT-LAMP assay will be helpful for detecting and preventing further SFTSV infection in China.

Background Real-time reverse transcription-PCR (rRT-PCR) has been the most effective and widely implemented diagnostic technology since the beginning of the COVID-19 pandemic. However, fuzzy rRT-PCR readouts with high Ct values are frequently encountered, resulting in uncertainty in diagnosis. Methods A Specific Enhancer for PCR-amplified Nucleic Acid (SENA) was developed based on the Cas12a trans -cleavage activity, which is specifically triggered by the rRT-PCR amplicons of the SARS-CoV-2 Orf1ab ( O ) and N fragments. SENA was first characterized to determine its sensitivity and specificity, using a systematic titration experiment with pure SARS-CoV-2 RNA standards, and was then verified in several hospitals, employing a couple of commercial rRT-PCR kits and testing various clinical specimens under different scenarios. Findings The ratio (10 min/5 min) of fluorescence change (FC) with mixed SENA reaction (mix- FCratio ) was defined for quantitative analysis of target O and N genes, and the Limit of Detection (LoD) of mix- FCratio with 95% confidence interval was 1.2≤1.6≤2.1. Totally, 295 clinical specimens were analyzed, among which 21 uncertain rRT-PCR cases as well as 4 false negative and 2 false positive samples were characterized by SENA and further verified by next-generation sequencing (NGS). The cut-off values for mix- FCratio were determined as 1.145 for positive and 1.068 for negative. Interpretation SENA increases both the sensitivity and the specificity of rRT-PCR, solving the uncertainty problem in COVID-19 diagnosis and thus providing a simple and low-cost companion diagnosis for combating the pandemic. Funding Detailed funding information is available at the end of the manuscript.

A multi-trait selective breeding program of Macrobrachium rosenbergii was initiated in China in 2015. In this program, the M . rosenbergii resources were widely collected from four countries, the origin of the founders was verified with 16 microsatellites and the pedigree was reconstructed, and the optimum contribution selection was used to make the mating design. In this study, we evaluated the genetic parameters and selection response for the harvest body weight (HBW) of M . rosenbergii after being communally reared for 95–109 days. The data were collected from two generations that comprised 25,212 progenies from 150 sires and 198 dams. The residual maximum-likelihood methodology was employed to evaluate the variance components, by fitting an animal model. The accuracy of estimated breeding values increased by 0.38% after pedigree reconstruction using microsatellite markers. The estimated heritability ( h 2 ) for HBW was moderate (0.212 ± 0.049) and the common environmental coefficient ( c 2 ) was low (0.063 ± 0.017) when all the data were used for the analysis. Within generations, h 2 was moderate to high (0.198 ± 0.080 to 0.338 ± 0.049). c 2 could only be estimated in G 1 , which was 0.055 ± 0.030. The average HBW of males was significantly larger than that of females ( P < 0.01). h 2 estimated for female HBWs were higher than that for males within generations, while h 2 estimated for female HBWs were lower than that for males across generations. But they were not significantly different ( P > 0.05). The genetic correlations between sexes were moderate to high within each generation (0.529 to 0.763). Two methods were used to estimate the realized response. One method was calculated from the differences between the least squares means of the selected population HBW and that of control population HBW, which was 14.01%. The other method was calculated from the differences between the EBVs of the selected population HBW and that of control population HBW, which was 11.52%. The predicted responses derived from two sets of genetic parameters acquired from within- and across- generation datasets were 11.68% and 10.67%, respectively. The present study provides valuable information for breeding programs of M . rosenbergii .

The iron prawn syndrome (IPS), also described as sexual precocity, results in the reduced size of farmed prawns at harvest and significant economic losses. IPS has been frequently reported in populations of farmed Macrobrachium rosenbergii since 2010, but the cause was heretofore unknown.

Comorbidities for fatal outcome among the COVID-19 patients: A hospital-based case-control study Dear Editor, Since the discovery of coronavirus disease 2019 (COVID-19), there have been numerous evidences supporting the pathogenic role of chronic comorbidities in the prognosis of infections, including the study by Galloway et al., 1 however, with the extent of the risk remained controversial. 2-6 The existing univariate models do not adequately distinguish between risk due to age and that due to increased presence of co-morbidities in older patients, thus these assessments of real effect from comorbidities are inevitably confounded. Here by performing a retrospective multi-center study, we try to evaluate the adjusted effect of the common preexisting comorbidities on COVID-19 related death, based on which, the therapy effect of three widely used anti-hypertension drugs were assessed. From 18 January to 24 February 2020, 1138 confirmed COVID-19 patients consisting of 920 survivals and 218 deaths from three designated hospitals for COVID-19 treatment in Hubei province were included for analysis (Table S1). The presence of comorbidities was reported in 49.12% (559/1138) of the total patients, with significantly higher frequency in the deceased than in the survivals (77.06% vs. 42.50%, P < 0.001; Table S1). As a whole, hypertension was the most prevalent comorbidity (32.95%), followed by diabetes mellitus (DM, 15.64%) and chronic heart diseases (CHD, 9.31%). The chronic obstructive pulmonary diseases (COPD), malignancy, cerebrovascular diseases (CVD), chronic kidney diseases (CKD), and chronic viral hepatitis (CVH) were less frequent, with prevalence ranging from 2.11% to 6.41%. By multivariate logistic regression model adjusting age, sex, and delay from symptom onset to hospital admission, six comorbidities showed significant associations with the disease outcome, with malignancy exhibiting the highest risk of death, followed by CKD, CVD, hypertension, CHD, and DM (Fig. 1 A and Table S2). An age-stratified analysis revealed the effect of comorbidities on death was reduced as the age increased. Among the patients ≤60 years, CVD had the highest effect on death, followed by hypertension. Among the patients aged 60-70 years, only malignancy and DM were related to fatal outcome. Among the patients aged > 70 years, none of the eight comorbidities demonstrated the significant association with fatal outcome. The sex-stratified analysis disclosed that male patients presenting with any of the three comorbidities (hypertension, DM, or CVD) had an increased risk of developing fatal outcome, in contrast, female patients presenting any of the four comorbidities (CHD, COPD, malignancy, or CKD) had an increased risk of fatal outcome (Table S3).