mNGS协助诊断无焦痂型丛林斑疹伤寒1例

 丛林斑疹伤寒是由恙虫病东方体感染引起的一种急性人畜共患传染病。临床表现为发热、皮疹、淋巴结肿大,被叮咬部位出现特异性焦痂和溃疡是该病诊断的最主要依据,没有焦痂和溃疡的患者往往很难识别。本文报告1例54岁男性患者,以发热、淋巴结肿大及全身多个脏器损伤为主要临床表现,因无特征性焦痂,且经验性使用抗菌药物效果不佳,给诊疗带来困难。病例标本送宏基因组二代测序(mNGS)明确病原体为恙虫病东方体。换用左氧氟沙星及阿奇霉素治疗后患者症状很快好转,最终痊愈出院。mNGS已被证明比传统培养方法更敏感,在早期诊断恙虫病方面优于传统临床方法,尤其是对于没有典型焦痂的患者。     

鹦鹉热衣原体肺炎患者临床特点分析

 目的 探讨分析鹦鹉热衣原体肺炎患者的临床表现及辅助检查结果特征,以提高对该病的认识。方法 回顾性分析某医院2019年6月- 2021年8月经宏基因组二代测序（mNGS）诊断鹦鹉热衣原体肺炎的9例患者临床资料。结果 9例患者中男性7例,女性2 例,包含1例妊娠36⁺²周孕妇;年龄25~70岁,中位年龄63岁;临床主要表现为高热（9例）、咳嗽（9例）、咳痰（7例）。白细胞计数正常8例,淋巴细胞计数降低9例,CRP升高9例,ESR升高9例,PCT正常4例,门冬氨酸氨基转移酶和丙氨酸氨基转移酶升高6例。9例患者胸部 CT均表现为斑片或斑片实变影。经血及肺泡灌洗液mNGS确诊后使用多西环素、莫西沙星、左氧氟沙星、阿奇霉素单药治疗方案,疗程14~21 d,均好转出院。结论 鹦鹉热衣原体肺炎临床表现多样、诊断困难,mNGS检测可以快速明确病原学诊断,有助于及时启动特异性抗感染治疗,可减少抗菌药物的使用,改善预后。     

PCR在猴B病毒鉴定中的应用研究

目的为鉴定新分离毒株是否为B病毒.方法根据ScinicarielloF报道的引物,用PCR方法扩增BV147、HSV-1、HSV-2,对扩增产物进行SacⅡ内切酶消化.结果这一对引物可同时对这3种病毒进行扩增,但只有BV147的扩增产物可被SacⅡ内切酶切开.对BV147扩增片段克隆测序的结果证实,其与美国B病毒E2490株部分基因(UL27)相对应位置的核苷酸同源性为100%.结论初步建立了检测B病毒DNA的PCR方法并测定了新分离病毒毒株的部分基因序列,证明新分离的病毒为B病毒.     

DNA直接银染测序法的建立和优化

目的：建立一种简便易行,灵敏经济的DNA直接银染测序法。方法：以标准DNApGEMR -3Zf( )(1g/L）为测序模板,测序电泳后,分别用银染试剂盒的银染色法及改良的银染色法进行染色,比较这两种方法的优劣。结果：改良的银染法具有操作简便,对显影温度不苛求,耗费少等优点。所显示的DNA带型与传统的方法比较,同样清晰可读,结论：优化的DNA直接银染测序法因简便经济,无放射性污染,更适合大多数实验室使用。     

Preliminary study of single nucleotide polymorphisms of PRPS2 gene in overproducing type of gouty patients

目的 :探讨编码人磷酸核糖焦磷酸合成酶亚单位 2的基因 PRPS2单核苷酸多态性与产生过剩型痛风患者的关系。方法 :利用聚合酶链反应扩增健康人和产生过剩型痛风患者 PRPS2基因全部外显 (包括外显子与内含子交界区 )的片段 ,采用多荧光标记的 PCR单链构象多态性分析技术对扩增的片段进行了筛选 ,对筛选到的片段进行序列测定 ,并与正常序列进行对照分析。结果 :在 PRPS2基因的第一个外显子区发现了一个 SNP( exon1+45A/G) ,第六个内含子区发现了一个 SNP ( intron6+12G/A) ,健康人与患者间的频率比较分别为 P =0 .0 96和 P =0 .2 73。结论 :提供了 PRPS2基因 SNP的数据库信息 ,为研究痛风发病机制提供了新的途径。     

Two-stage Study of Familial Prostate Cancer by Whole-exome Sequencing and Custom Capture Identifies 10 Novel Genes Associated with the Risk of Prostate Cancer

BackgroundFamily history of prostate cancer (PCa) is a well-known risk factor, and both common and rare genetic variants are associated with the disease.ObjectiveTo detect new genetic variants associated with PCa, capitalizing on the role of family history and more aggressive PCa.Design, setting, and participantsA two-stage design was used. In stage one, whole-exome sequencing was used to identify potential risk alleles among affected men with a strong family history of disease or with more aggressive disease (491 cases and 429 controls). Aggressive disease was based on a sum of scores for Gleason score, node status, metastasis, tumor stage, prostate-specific antigen at diagnosis, systemic recurrence, and time to PCa death. Genes identified in stage one were screened in stage two using a custom-capture design in an independent set of 2917 cases and 1899 controls.Outcome measurements and statistical analysisFrequencies of genetic variants (singly or jointly in a gene) were compared between cases and controls.Results and limitationsEleven genes previously reported to be associated with PCa were detected (ATM, BRCA2, HOXB13, FAM111A, EMSY, HNF1B, KLK3, MSMB, PCAT1, PRSS3, and TERT), as well as an additional 10 novel genes (PABPC1, QK1, FAM114A1, MUC6, MYCBP2, RAPGEF4, RNASEH2B, ULK4, XPO7, and THAP3). Of these 10 novel genes, all but PABPC1 and ULK4 were primarily associated with the risk of aggressive PCa.ConclusionsOur approach demonstrates the advantage of gene sequencing in the search for genetic variants associated with PCa and the benefits of sampling patients with a strong family history of disease or an aggressive form of disease.Patient summaryMultiple genes are associated with prostate cancer (PCa) among men with a strong family history of this disease or among men with an aggressive form of PCa.     

A novel SRD5A2 mutation in an Iranian family with sex development disorder

Disorders of sex development (DSD) are different types of conditions that their accurate diagnosis by using conventional phenotypic and biochemical approaches is a challenging issue. Precise determination of DSD is critical due to the detection of possible life-threatening associated disorders. It may also assist parents in choosing the most suitable management for their affected child. In this study, two affected kids born from consanguineous families who were clinically diagnosed for sex development disorder were investigated for the main cause of the disease. Biochemical analysis failed to make an accurate diagnosis. Karyotype analysis showed an abnormal sex chromosome pattern. Whole exome sequencing was sequentially applied to precisely ascertain the genetic cause of the disease. A novel deletion, g.40936_53878del12943insTG (NG_008365.1), and one known mutation, c.586G>A (p.Gly196Ser), were detected in SRD5A2 gene in case I and case II respectively. Further analysis was performed using polymerase chain reaction, primer walking and Sanger sequencing to detect the nucleotides changes accurately. Segregation analysis in the families confirmed 13kb novel homozygous deletion of SRD5A2 in case I and c.586G>A in case II. The present study confirms the diagnostic value of whole exome sequencing in the detection of DSD aetiology, especially when several differential diagnoses are possible.     

基于高通量测序的人工培殖冬虫夏草僵虫颜色变化转录组分析

目的:探索人工培殖冬虫夏草Cordyceps sinensis(BerK.)Sacc.外观品质形成的分子基础。方法:以重庆和康定人工培殖冬虫夏草僵虫为材料,利用Illumina Hiseq 2500高通量测序技术进行转录组测序分析,基于差异倍数(FC)和错误发现率(FDR)筛选差异表达基因,具体筛选条件为|log2FC|>1、FDR<0.05。结果:共获得21942个转录本,利用DEGSeq筛选获得1575个差异表达基因,其中上调表达基因626个、下调表达基因949个;基因本体(GO)数据库和京都基因与基因组百科全书(KEGG)富集分析结果显示,这些差异表达基因主要集中于氨基酸代谢、折叠-分选-降解、糖代谢、转运和分解代谢、辅酶因子和微生物代谢等途径;筛选到66个颜色变化差异表达基因,包含46个氧化还原酶活性相关基因、12个过氧化物酶体相关基因、3个单加氧酶活性调节相关基因、3个黑化反应相关基因、2个活性氧代谢相关基因,其中tyr1、TYR基因可能是冬虫夏草僵虫颜色变化的关键功能基因。结论:利用高通量测序技术和生物信息分析获得了人工培殖冬虫夏草僵虫转录组信息特征,为阐明人工培殖冬虫夏草僵虫颜色变化的调控机制研究提供参考。