1.Establishment and evaluation of a genome-wide multiplex PCR sequencing method for severe fever with thrombocytopenia syndrome virus based on high-throughput sequencing technology
Xiaoyang WANG ; Shujun PEI ; Zeqian CHEN ; Wenshuai SUO ; Zhijie YI ; Yi LI ; Xueyong HUANG ; Wanshen GUO
Chinese Journal of Microbiology and Immunology 2022;42(12):931-939
Objective:To establish a sequencing method for the genome of severe fever with thrombocytopenia syndrome virus (SFTSV) based on next-generation sequencing (NGS).Methods:SFTSV RNA was extracted from serum samples of patients with severe fever with thrombocytopenia syndrome. SFTSV-specific primers were designed using Primer 5.0 software. A multiplex PCR method was constructed and used to amplify the nucleotide sequence of SFTSV. Whole-genome sequencing was performed on the NGS platform.Results:The whole genes of SFTSV isolates in 28 serum samples were amplified by the multiplex PCR with a coverage over 94%. Sequencing and phylogenetic analysis of those strains revealed that the predominant strains ( n=20) belonged to genotype A, followed by genotypes B ( n=4) and E ( n=3). Conclusions:A high-throughput sequencing method for SFTSV based on multiplex PCR was established in this study. This method was characterized by high specificity and good quality and could improve the sequencing efficiency.
2.Epidemic characteristics and spatiotemporal distribution analysis of human brucellosis in Henan Province, 2005 - 2021
Wenshuai SUO ; Shujun PEI ; Xiaoyang WANG ; Zeqian CHEN ; Zhijie YI ; Jiarui LYU ; Baifan ZHANG ; Ying YE ; Xueyong HUANG
Chinese Journal of Endemiology 2023;42(7):540-547
Objective:To analyze the epidemiological characteristics and spatiotemporal characteristics of human brucellosis in Henan Province.Methods:Data of human brucellosis in Henan Province from 2005 to 2021 were collected through the China Disease Prevention and Control Information System, and a descriptive epidemiological method was used to analyze the epidemic profile of brucellosis in Henan Province and the three distribution characteristics. Global and local spatial autocorrelation were used to analyze the spatial distribution and the hot spots of brucellosis in Henan Province, respectively, and spatiotemporal scanning was used to analyze the spatiotemporal clustering regions of brucellosis in Henan Province.Results:A total of 39 862 brucellosis cases were reported in Henan Province from 2005 to 2021, with an average annual incidence of 2.44/100 000, and the number of cases showed an overall increasing trend each year (χ 2trend = 11 127.85, P = 0.001). The onset months were mainly concentrated from March to July, accounting for 59.00% (23 517/39 862), with May as the peak (5 478 cases). Cases of brucellosis were reported in 157 counties (cities, districts) of the province. The ratio of male to female was 2.52∶1.00 (28 542/11 320). Farmers were the main occupation, with 32 985 cases (82.75%). The age of onset was mainly 45 to 65 years old, with 20 226 cases (50.74%). The global spatial autocorrelation analysis showed that the global Moran's I was > 0, Z > 1.96, and P < 0.05 in all years except 2006 - 2008, showing spatial clustering. Further local spatial autocorrelation analysis was performed, and high-high and low-low clustering areas existed in 2012 - 2021 ( P < 0.01). Spatiotemporal scanning analysis showed that there was one spatiotemporal cluster in the high incidence area and two spatiotemporal clusters in the low incidence area. The high incidence cluster was centered in Neixiang County, covering 48 counties (cities, districts) including Song County and Ruzhou City, and the aggregation time was from 2014 to 2021. The two low incidence clusters were centered in Yongcheng City and Boai County, covering 58 and 18 counties (cities, districts), respectively, and the aggregation time was 2016 - 2021 and 2005 - 2012, respectively. Conclusion:The overall incidence of brucellosis in Henan Province is on the rise from 2005 to 2021, with middle-aged and elderly male farmers as the main affected population, and there are spatiotemporal clusters of brucellosis in Henan Province.
3.Epidemiological investigation and genomic characterization of the first case of fever with thrombocytopenia syndrome in Puyang
Zeqian CHEN ; Yi LI ; Xiaoyang WANG ; Shujun PEI ; Wenshuai SUO ; Zhiquan HE ; Aiguo YOU ; Yibin HAO ; Xueyong HUANG
Chinese Journal of Microbiology and Immunology 2023;43(8):627-633
Objective:To report the first case of sever fever with thrombocytopenia syndrome caused by severe fever with thrombocytopenia syndrome virus (SFTSV) in Puyang city, and to study the epidemiological and molecular characteristics of S, M, L fragments of the SFTSV isolate.Methods:The epidemiological characteristics of this case was analyzed with epidemiological methods. SFTSV was isolated from the patient′s serum sample. Nucleic acid of SFTSV was extracted and detected by fluorescent RT-PCR. A multiplex PCR method was constructed to amplify the nucleic acid sequence of the virus. whole-genome sequencing was performed on the next-generation sequencing platform. MEGA11 and DNAStar was used for homology analysis and a phylogenetic tree was constructed.Results:Epidemiological investigation showed that the patient and his close contacts had no history of travel or tick bite within 14 d, but had a history of fieldwork. The patient′s serum sample was positive for SFTSV nucleic acid. Genetic analysis showed that the S, M, L gene fragments of the first SFTSV isolate in Puyang belonged to genotype E. This isolate shared 94.8%-99.6%, 94.0%-99.8% and 95.7%-99.7% nucleotide sequence homology with the representative strains acquired from GeneBank in S, M, L gene fragments, respectively.Conclusions:This case was the first case of SFTSV-caused severe fever with thrombocytopenia syndrome in Puyang. The SFTSV isolate shared a close homology with domestic isolates, but its genotype was significantly different from the SFTSV strains isolated in Henan in recent years, indicating that it might an imported case from other places in Henan Province or Hubei Province. Disease monitoring and professional training for medical personnel should be strengthened and more attention should be paid to the evolution and mutation of SFTSV.
4.Study on Traditional Chinese Medicine Syndrome Characteristics of 1676 Heart Failure Inpatients: A Cross-Sectional Survey Based on Real-World Electronic Medical Record Information
Yi DU ; Zheng LI ; Guanlin YANG ; Shuqi DONG ; Wenshuai HUANG ; Nanxing XIAN ; Puyu GUO ; Jiajie QI ; Bohang CHEN ; Xin XU ; Zhe ZHANG ; Yi YANG
Journal of Traditional Chinese Medicine 2024;65(3):299-307
ObjectiveTo analyse the clinical characteristics of different traditional Chinese medicine (TCM) syndromes in patients with heart failure based on information from electronic medical record. MethodsA cross-sectional study was conducted to collect clinical data of all inpatients with heart failure in the Department of Cardiology, Affiliated Hospital of Liaoning University of Traditional Chinese Medicine from January 1, 2019 to December 31, 2020. A database of clinical TCM data was established to explore the characteristics of clinical data of basic information, syndromes and syndrome element types, and biochemical indexes. The distribution of TCM syndromes and syndrome elements in heart failure patients were also analysed, and the basic information and biochemical indexes of the patients with top 7 different TCM syndrome types were compared. ResultsA total of 1676 inpatients with heart fai-lure were included. The top 7 TCM syndromes of heart failure were syndrome of phlegm turbidity and blood stasis (477 cases, 28.46%), syndrome of qi deficiency and blood stasis (439 cases, 26.19%), syndrome of qi deficiency and blood stasis with water retention (274 cases, 16.35%), syndrome of yang deficiency with water retention (145 cases, 8.65%), syndrome of qi and yin deficiency (104 cases, 6.21%), syndrome of qi and yin deficiency with blood stasis (80 cases, 4.77%), syndrome of heart yang deficiency (59 cases, 3.52%). Among the 1676 patients, 6 syndrome elements accounted for more than 5%. Blood stasis accounted for the highest proportion of TCM syndrome element type (1292 cases, 77.09%), followed by qi deficiency (919 cases, 54.83%), phlegm (498 cases, 29.71%), water retention (434 cases, 25.89%), yang deficiency (215 cases, 12.82%) and yin deficiency (191 cases, 11.40%). Among the 1676 patients, 1308 cases of acute heart failure mainly showed syndrome of phlegm turbidity and blood stasis (386 cases, 29.51%), and 368 of chronic heart fai-lure mainly showed syndrome of qi deficiency and blood stasis (118 cases, 32.07%). Patients with syndrome of phlegm turbidity and blood stasis had the shortest disease duration of 0.3 months, while those with syndrome of heart yang deficiency had the longest disease duration of 15 months. The proportion of syndrome of phlegm turbidity and blood stasis was the highest in patients with heart failure combined with coronary artery disease, the proportion of syndrome of qi deficiency and blood stasis with water retention was the highest in patients with heart failure combined with atrial fibrillation, and the proportion of patients with syndrome of qi deficiency and blood stasis with water retention and syndrome of yang deficiency with water retention in those applying diuretics during the hospital stay was the highest with more than 86%. The different 7 TCM syndromes showed statistically difference in patients with complications including coronary artery disease, old myocardial infarction, atrial fibrillation, pre and post-admission medication usage including intravenous vasodilators, cardiac stimulants, diuretics, and level of blood chloride, blood urea, blood creatinine, blood bicarbonate, blood albumin, and blood total bilirubin (P<0.05). ConclusionThe most common TCM syndromes in patients with heart failure are syndrome of phlegm turbidity and blood stasis and syndrome of qi deficiency and blood stasis. Different TCM syndromes have different characteristics in gender, disease complications, medication before and after admission, and blood indexes.