Inferring Mycobacterium Tuberculosis Drug Resistance and Transmission using Whole-genome Sequencing in a High TB-burden Setting in China

Objective China is among the 30 countries with a high burden of tuberculosis(TB)worldwide,and TB remains a public health concern.Kashgar Prefecture in the southern Xinjiang Autonomous Region is considered as one of the highest TB burden regions in China.However,molecular epidemiological studies of Kashgar are lacking.Methods A population-based retrospective study was conducted using whole-genome sequencing(WGS)to determine the characteristics of drug resistance and the transmission patterns.Results A total of 1,668 isolates collected in 2020 were classified into lineages 2(46.0%),3(27.5%),and 4(26.5%).The drug resistance rates revealed by WGS showed that the top three drugs in terms of the resistance rate were isoniazid(7.4%,124/1,668),streptomycin(6.0%,100/1,668),and rifampicin(3.3%,55/1,668).The rate of rifampicin resistance was 1.8%(23/1,290)in the new cases and 9.4%(32/340)in the previously treated cases.Known resistance mutations were detected more frequently in lineage 2 strains than in lineage 3 or 4 strains,respectively:18.6%vs.8.7 or 9%,P<0.001.The estimated proportion of recent transmissions was 25.9%(432/1,668).Multivariate logistic analyses indicated that sex,age,occupation,lineage,and drug resistance were the risk factors for recent transmission.Despite the low rate of drug resistance,drug-resistant strains had a higher risk of recent transmission than the susceptible strains(adjusted odds ratio,1.414;95%CI,1.023–1.954;P=0.036).Among all patients with drug-resistant tuberculosis(DR-TB),78.4%(171/218)were attributed to the transmission of DR-TB strains.Conclusion Our results suggest that drug-resistant strains are more transmissible than susceptible strains and that transmission is the major driving force of the current DR-TB epidemic in Kashgar.

Genomic surveillance of emerging SARS-CoV-2 Omicron variations in Tianjin Municipality, China 2022

The coronavirus disease 2019(COvID-19)pandemic caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)has severely impacted public health.In 2022,the Omicron variant of SARS-CoV-2 rapidly became the dominant circulating variant in the local COVID-19 outbreaks in Tianjin Municipality,China.To gain a deeper understanding of the genetic variations of the Omicron variant in Tianjin,specimens from indi-viduals who tested positive for SARS-CoV-2 between December 2021 and November 2022 were used for virus whole genome sequencing and phylogenetic analysis.A total of 1,674 high-quality Omicron sequences were obtained,consisting of 1,339 sequences from local cases belonging to 20 Phylogenetic Assignment of Named Global Outbreak(PANGO)lineages and 335 sequences from imported cases belonging to 70 lineages.Tianjin experienced five waves of local outbreaks,accompanied by multiple substitutions among subvariants,ranging from the initial BA.1.1 lineage to the subsequent BA.2,BF.7,and BA.5.2 lineages.The evolutionary rate of local strains,estimated to be 28.999 substitutions per year,and the evolutionary rate of imported strains,estimated to be 24.946 substitutions per year,were lower than that of the strains circulating globally.The addi-tional substitutions and deletions of local strains have been used to identify and disrupt the virus transmission chains.The subvariants such as BA.5.2.48,BA.5.2.49,BF.7.14,and XBB.1 circulating in the fifth epidemic wave presented criterial immune escape mutations including S:R346T,S:L452R and S:F486V.It is essential to implement genomic surveillance strategies to investigate further the development of genomic mutation char-acteristics in the SARS-CoV-2 variant.This ongoing monitoring will contribute to a better understanding of the virus's genetic changes and aid in effective control measures.

The mutational dynamics of the SARS-CoV-2 virus in serial passages in vitro

Since its outbreak in 2019,Severe Acute Respiratory Syndrome Coronavirus 2(SARS-Co V-2)keeps surprising the medical community by evolving diverse immune escape mutations in a rapid and effective manner.To gain deeper insight into mutation frequency and dynamics,we isolated ten ancestral strains of SARS-Co V-2 and performed consecutive serial incubation in ten replications in a suitable and common cell line and subsequently analysed them using RT-q PCR and whole genome sequencing.Along those lines we hoped to gain fundamental insights into the evolutionary capacity of SARS-Co V-2 in vitro.Our results identified a series of adaptive genetic changes,ranging from unique convergent substitutional mutations and hitherto undescribed insertions.The region coding for spike proved to be a mutational hotspot,evolving a number of mutational changes including the already known substitutions at positions S:484 and S:501.We discussed the evolution of all specific adaptations as well as possible reasons for the seemingly inhomogeneous potential of SARS-Co V-2 in the adaptation to cell culture.The combination of serial passage in vitro with whole genome sequencing uncovers the immense mutational potential of some SARS-Co V-2 strains.The observed genetic changes of SARS-Co V-2 in vitro could not be explained solely by selectively neutral mutations but possibly resulted from the action of directional selection accumulating favourable genetic changes in the evolving variants,along the path of increasing potency of the strain.Competition among a high number of quasi-species in the SARS-Co V-2 in vitro population gene pool may reinforce directional selection and boost the speed of evolutionary change.