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| 1 | 心肌酶谱和高敏肌钙蛋白T在小儿手足口病并发心肌损伤诊断中的临床应用价值显示文摘目的探析心肌酶谱、高敏肌钙蛋白T诊断小儿手足口病并发心肌损伤的临床应用价值。方法选择2016年4月至2017年1月湖北医药学院附属太和医院收治的手足口病患儿100例,根据心电图检查结果分为心电图正常组(n=40)和心电图异常组(n=60),同期选择体检健康者50例为对照组,比较三组的心肌酶同工酶(CK-MB)、肌酸激酶(CK)、α-羟丁酸脱氢酶(α-HBDH)、乳酸脱氢酶(LDH)、天冬氨酸氨基转移酶(AST)、高敏肌钙蛋白T(hs-cTnT),并比较手足口病异常心电图组患儿急性期与恢复期的hs-cTnT、CK等生化指标。结果手足口病患儿心电图异常组的CK-MB、CK、α-HBDH、LDH、AST、hs-cTnT明显高于手足口病患儿心电图正常组和对照组,差异有统计学意义(P<0.05)。手足口病患儿心电图异常组急性期CK-MB、CK、α-HBDH、LDH、AST、hs-cTnT水平明显高于手足口病患儿心电图异常组恢复期,差异均有统计学意义(P<0.05)。结论心肌损伤为手足口病患儿的严重并发症,监测患儿的hs-cTnT、心肌酶谱等生化指标对手足口病患儿心肌损伤的诊治具有重要的临床价值。 | 付红霞 杨靖 徐祥 | 2018 | 国际检验医学杂志2018，39，8： | 13 |
| 2 | TaqMan Real-time RT-PCR Assay for Detecting and Differentiating Japanese Encephalitis Virus显示文摘Objective To detect Japanese encephalitis virus(JEV) rapidly and distinguish its genotypes, a TaqMan-based reverse transcriptase quantitative polymerase chain reaction(RT-PCR) detection system was developed.Methods By aligning the full-length sequences of JEV(G1-G5), six sets of highly specific TaqMan real-time RT-PCR primers and probes were designed based on the highly conserved NS1, NS2, and M genes of JEV, which included one set for non-specific JEV detection and five sets for the detection of specific JEV genotypes. Twenty batches of mosquito samples were used to evaluate our quantitative PCR assay.Results With the specific assay, no other flavivirus were detected. The lower limits of detection of the system were 1 pfu/mL for JEV titers and 100 RNA copies/μL. The coefficients of variation of this real-time RT-PCR were all < 2.8%. The amplification efficiency of this method was between 90% and 103%.Conclusion A TaqMan real-time RT-PCR detection system was successfully established to detect and differentiate all five JEV genotypes. | SHAO Nan LI Fan NIE Kai FU Shi Hong ZHANG Wei Jia HE Ying LEI Wen Wen WANG Qian Ying LIANG Guo Dong CAO Yu Xi WANG Huan Yu | 2018 | Biomedical and Environmental Sciences2018，31，3： | 10 |
| 3 | 手足口病实验室检测方法的研究进展显示文摘手足口病(hand-foot-and-mouth disease,HFMD)是由多种人肠道病毒引起的儿童常见传染病,以发热和手、足、口腔等部位的皮疹或疱疹为主要症状,少数患者出现严重的神经后遗症,个别重症患儿病情进展快可导致死亡。了解手足口病病毒和可能与疾病发展相关病毒的病原学检测方法有利于进一步了解手足口病的传播模式和分子流行病学研究进展,有助于临床的对症治疗及预防控制该疾病的蔓延。本文就手足口病实验室检测方法的研究进展进行综述,为一线工作人员在临床实践过程中选择合适的HFMD检测方法提供参考。 | 欧晓燕 许彬 | 2018 | 分子诊断与治疗杂志2018，10，6： | 7 |
| 4 | A Reverse-transcription Recombinase-aided Amplification Assay for the Rapid Detection of the Far-Eastern Subtype of Tick-borne Encephalitis Virus显示文摘Objective Tick-borne encephalitis virus(TBEV) is an emerging pathogen in Europe and North Asia that causes tick-borne encephalitis(TBE). A simple, rapid method for detecting TBEV RNA is needed to control this disease. Methods A reverse-transcription recombinase-aided amplification(RT-RAA) assay was developed. This assay can be completed in one closed tube at 39℃ within 30 minutes. The sensitivity and specificity of RT-RAA were validated using non-infectious synthetic RNA representing a fragment of the NS5 region of the wild-type(WT) TBEV genome and the Senzhang strain. Additionally, 10 batches of tick samples were used to evaluate the performance of the RT-RAA assay. Results The analytical limit of detection of the assay was 20 copies per reaction of the TBEV synthetic transcript and 3 plaque-forming units(pfu) per reaction of TBEV titers. With the specific assay, no signal due to other arboviruses was observed. Of the 10 batches of tick samples obtained from the Changbai Mountains of China, three were TBEV-positive, which was consistent with the results of the quantitative real-time PCR assay. Conclusion A rapid, highly sensitive, specific, and easy-to-use method was developed for the detection of the TBEV Far-Eastern subtype. | WANG Qian Ying LI Fan SHEN Xin Xin FU Shi Hong HE Ying LEI Wen Wen LIANG Guo Dong WANG Huan Yun MA Xue Jun | 2019 | Biomedical and Environmental Sciences2019，32，5： | 4 |
| 5 | Identification of Histoplasma causing an unexplained disease cluster in Matthews Ridge, Guyana显示文摘Here,we report the identification of Histoplasma causing an unexplained disease cluster in Matthews Ridge,Guyana.In March 2019,14 employees of Chongqing Bosai Mining Company,China,working in a manganese mining of Guyana,had unexplained fever,and two of them died.We obtained lung and brain tissues as well as the blood samples from the two deceased cases(patient No.1 and 2),and bronchoscopy lavages and cerebrospinal fluid samples from one severe case(patient No.3),respectively.All samples were tested by pathological examination,high-throughput sequencing,and real-time PCR.Pathological detection showed the presence of spore-like structures in the lung tissue of patient No.1,indicating a fungal infection in this patient.Nanopore sequencing identified the existing of H.capsulatum in the lung tissue sample within 13 h.Next-generation sequencing identified specific fragments of H.capsulatum in all of the samples tested(lung,brain and blood serum from the deceased cases,and plasma from the severe case).Real-time PCR assays did not reveal any viral infection related to transmission from bat feces.We conclude that H.capsulatum was the causative pathogen of this disease cluster based on epidemiologic,clinical,pathological and nucleic acid evidence. | Ji Wang Weimin Zhou Hua Ling Xiaoping Dong Yi Zhang Jiandong Li Yong Zhang Jingdong Song William J.Liu Yang Li Ruiqing Zhang Wei Zhen Kun Cai Shuangli Zhu Dongyan Wang Jinbo Xiao Yigang Tong Wenli Liu Lihua Song Wei Wu Yang Liu Xiang Zhao Ruihuan Wang Sheng Ye Jing Wang Roujian Lu Baoying Huang Fei Ye Wenwen Lei Rongbao Gao Qi Shi Cao Chen Jun Han Wenbo Xu George F.Gao Xuejun Ma Guizhen Wu | 2019 | Biosafety and Health2019，1，3： | 3 |
| 6 | VSITA, an Improved Approach of Target Amplification in the Identification of Viral Pathogens显示文摘Objective Unbiased next generation sequencing(NGS) is susceptible to interference from host or environmental sequences. Consequently, background depletion and virome enrichment techniques are usually needed for clinical samples where viral load is much lower than background sequences. Methods A viral Sequence Independent Targeted Amplification(VSITA) approach using a set of non-ribosomal and virus-enriched octamers(V8) was developed and compared with traditionally used random hexamers(N6). Forty-five archived clinical samples of different types were used in parallel to compare the V8 and N6 enrichment performance of viral sequences and removal performance of ribosomal sequences in the step of reverse transcription followed by quantitative PCR(qP CR). Ten sera samples from patients with fever of unknown origin and 10 feces samples from patients with diarrhea of unknown origin were used in comparison of V8 and N6 enrichment performance following NGS analysis. Results A minimum 30 hexamers matching to viral reference sequences(sense and antisense) were selected from a dataset of random 4,096(4~6) hexamers(N6). Two random nucleotides were added to the 5' end of the selected hexamers, and 480(30 × 4~2) octamers(V8) were obtained. In general, VSITA approach showed higher enrichment of virus-targeted c DNA and enhanced ability to remove unwanted ribosomal sequences in the majorities of 45 predefined clinical samples. Moreover, VSITA combined with NGS enabled to detect not only more viruses but also achieve more viral reads hit and higher viral genome coverage in 20 clinical samples with diarrhea or fever of unknown origin. Conclusion The VSITA approach designed in this study is demonstrated to possess higher sensitivity and broader genome coverage than traditionally used random hexamers in the NGS-based identification of viral pathogens directly from clinical samples. | ZHANG Yi ZHANG Chen LI Bo LI Yang HE Xiao Zhou LI Acher WU Wei DUAN Su Xia QIU Fang Zhou WANG Ji SHEN Xin Xin YANG Meng Jie LI De Xin MA Xue Jun | 2018 | Biomedical and Environmental Sciences2018，31，4： | 3 |
| 7 | Next-generation Sequencing Study of Pathogens in Serum from Patients with Febrile Jaundice in Sierra Leone显示文摘Objective People in Western Africa suffer greatly from febrile jaundice, which is caused by a variety of pathogens. However, yellow fever virus(YFV) is the only pathogen under surveillance in Sierra Leone owing to the undeveloped medical and public health system there. Most of the results of YFV identification are negative. Elucidation of the pathogen spectrum is required to reduce the prevalence of febrile jaundice. Methods In the present study, we used Ion Torrent semiconductor sequencing to profile the pathogen spectrum in archived YFV‐negative sera from 96 patients in Sierra Leone who presented with unexplained febrile jaundice. Results The most frequently identified sequencing reads belonged to the following pathogens: cytomegalovirus(89.58%), Epstein‐Barr virus(55.21%), hepatitis C virus(34.38%), rhinovirus(28.13%), hepatitis A virus(20.83%), coxsackievirus(10.42%), Ebola virus(8.33%), hepatitis E virus(8.33%), lyssavirus(4.17%), leptospirosis(4.17%), chikungunya virus(2.08%), Crimean‐Congo hemorrhagic fever virus(1.04%), and hepatitis B virus(1.04%). Conclusion The distribution of sequencing reads suggests a broader spectrum of pathogens for consideration in clinical diagnostics and epidemiological surveillance in Sierra Leone. | ZHANG Yi YE Fei XIA Lian Xu ZHU Ling Wei IDRISSA Laybohr Kamara HUANG Ke Qiang ZHANG Yong LIU Jun BRIMA Kargbo WANG Ji LIANG Mi Fang SONG Jing Dong MA Xue Jun WU Gui Zhen | 2019 | Biomedical and Environmental Sciences2019，32，5： | 2 |
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