A Wax Interface-Enabled One-Pot Multiplexed Nucleic Acid Testing Platform for Rapid and Sensitive Detection of Viruses and Variants.

High-quality, low-cost, and rapid detection is essential for the society to reopen the economy during the critical period of transition from Coronavirus Disease 2019 (COVID-19) pandemic response to pandemic control. In addition to performing sustainable and target-driven tracking of SARS-CoV-2, conducting comprehensive surveillance of variants and multiple respiratory pathogens is also critical due to the frequency of reinfections, mutation immune escape, and the growing prevalence of the cocirculation of multiple viruses. By utilizing a 0.05 cents wax interface, a Stable Interface assisted Multiplex Pathogenesis Locating Estimation in Onepot (SIMPLEone) using nested RPA and CRISPR/Cas12a enzymatic reporting system is successfully developed. This smartphone-based SIMPLEone system achieves highly sensitive one-pot detection of SARS-CoV-2 and its variants, or multiple respiratory viruses, in 40 min. A total of 89 clinical samples, 14 environmental samples, and 20 cat swab samples are analyzed by SIMPLEone, demonstrating its excellent sensitivity (3-6 copies/reaction for non-extraction detection of swab and 100-150 copies/mL for RNA extraction-based assay), accuracy (>97.7%), and specificity (100%). Furthermore, a high percentage (44.2%) of co-infection cases are detected in SARS-CoV-2-infected patients using SIMPLEone's multiplex detection capability.

Sensitive and Rapid Diagnosis of Respiratory Virus Coinfection Using a Microfluidic Chip-Powered CRISPR/Cas12a System.

The ongoing pandemic caused by severe acute respiratory syndrome coronavirus 2 is profoundly influencing the global healthcare system and people's daily lives. The high resource consumption of coronavirus disease 2019 (COVID-19) is resulting in insufficient surveillance of coinfection or resurgence of other critical respiratory epidemics, which is of public concern. To facilitate evaluation of the current coinfection situation, a microfluidic system (MAPnavi) is developed for the rapid (<40 min) and sensitive diagnosis of multiple respiratory viruses from swab samples in a fully sealed and automated manner, in which a nested-recombinase polymerase amplification and the CRISPR-based amplification system is first proposed to ensure the sensitivity and specificity. This novel system has a remarkably low limit of detection (50-200 copies mL-1 ) and is successfully applied to detect 171 clinical samples (98.5% positive predictive agreement; 100% negative predictive agreement), and the results identify 45.6% coinfection among clinical samples from patients with COVID-19. This approach has the potential to shift diagnostic and surveillance efforts from targeted testing for a high-priority virus to comprehensive testing of multiple virus sets and to greatly benefit the implementation of decentralized testing.

[Driving Factors of the Dynamics of Microbial Community in a Dam of Copper Mine Tailings].

The relative importance of the deterministic versus stochastic processes underlying community dynamics has long been a central theme in community ecology, and is intensively debated in the field. Microbial communities play key roles in nutrient cycling and the flow of energy in ecosystems. The research on the structural dynamics of microbial community will provide data and theoretical support for understanding the assembly mechanisms of community, and for predicting the dynamics of microbial community under environmental stress. In this study, the Illumina MiSeq method was applied to investigate the structural dynamics of bacterial and fungal community in a dam of Shibahe mine tailings at different restoration stages (1-45 years). The results indicated that the soil physicochemical properties in the dam of mine tailings formed an ecological gradient, and the plant community showed succession along the restoration time. The diversity of plant communities was significantly correlated with soil nutrient contents but not with soil heavy metal contents. The structure of the microbial communities showed significant differences at different restoration stages of the dam land, in which Proteobacteria, Actinobacteria, Firmicutes, and Acidobacteria were the dominant bacterial phyla, and Ascomycota, Basidiomycota, and Zygomycota were the dominant fungal phyla. The assembly of the microbial community was shaped mainly by the soil nutrients and soil heavy metal contents, but plant diversity had no significant effect on the microbial community structure. It was suggested that edaphic factors drive the dynamics of microbial communities under the stress conditions of pH and heavy metals on small, local scales.

Ecological patterns and adaptability of bacterial communities in alkaline copper mine drainage.

Environmental gradient have strong effects on community assembly processes. In order to reveal the effects of alkaline mine drainage (AlkMD) on bacterial and denitrifying bacterial community compositions and diversity in tailings reservoir, here we conducted an experiment to examine all and core bacterial taxa and denitrifying functional genes's (nirS, nirK, nosZΙ) abundance along a chemical gradient in tailings water in Shibahe copper tailings in Zhongtiaoshan, China. Differences in bacterial and denitrifying bacterial community compositions in different habitats and their relationships with environmental parameters were analyzed. The results showed that the richness and diversity of bacterial community in downstream seeping water (SDSW) were the largest, while that in upstream tailings water (STW1) were the lowest. The diversity and abundance of bacterial communities tended to increase from STW1 to SDSW. The variation of bacterial community diversity was significantly related to electroconductibility (EC), nitrate (NO3-), nitrite (NO2-), total carbon (TC), inorganic carbon (IC) and sulfate (SO42-), but was not correlated with geographic distance in local scale. Core taxa from class to genus were all significantly related to NO3- and NO2-. Core taxa Rhodobacteraceae, Rhodobacter, Acinetobacter and Hydrogenophaga were typical denitrifying bacteria. The variation trends of these groups were consistent with the copy number of nirS, nirK and nosZΙ, demonstrating their importance in the process of nitrogen reduction. The copy number of nirK, nosZΙ and nirS/16S rDNA, nirK/16Sr DNA correlated strongly with NO3-, NO2- and IC, but nirS and nosZI/16SrDNA had no significant correlation with NO3- and NO2-. The copy numbers of denitrifying functional genes (nirS, nirK and nosZΙ) were negatively correlated with heavy metal plumbum (Pb) and zinc (Zn). It showed that heavy metal contamination was an important factor affecting the structure of denitrifying bacterial community in AlkMD. In this study we have identified the distribution pattern of bacterial community along physiochemical gradients in alkaline tailings reservoir and displayed the driving force of shaping the structure of bacterial community. The influence of NO3-, NO2-, IC and heavy metal Pb and Zn on bacterial community might via their influence on the functional groups involving nitrogen, carbon and metal metabolisms.

[Composition and Environmental Adaptation of Microbial Community in Shibahe Copper Tailing in Zhongtiao Mountain in Shanxi].

In order to reveal the effects of heavy metal pollution on microbial community compositions and microbial community diversity in tailing area,we conducted an experiment by examining the microbial community in tailing water,sediments and tailing sand in Shibahe copper tailing in Zhongtiao Mountain.Differences in microbial community compositions in different habitats and their relationships with environmental parameters were analyzed.The results showed that the richness and diversity of microbial community were the largest in tailing sand,but the lowest in tailing water.Microbial community compositions were similar between tailing water and sediments.There were significant positive correlations between the relative abundance of the dominant family (Sphingomonadaceae) and contents of heavy metals (Cd,Cu,Mn,Ni,Pb,Zn),while there were significant negative correlations between relative abundances of aulobacteraceae, Methylobacteriaceae, Nocardioidaceae, Microbacteriaceae, Micrococcaceae, Streptococcaceae and Paenibacillaceae and heavy metal contents.It showed that most of the bacteria were inhibited by heavy metals,but Sphingomonadaceae had a higher tolerance to heavy metals which may indicate that it has a potential for remediation of heavy metal contamination.