基因芯片技术检测结核杆菌耐药的临床研究

目的 评价基因芯片技术在结核病防治工作中对结核分枝杆菌异烟肼和利福平耐药性的检测效果.方法 利用基因芯片技术对38例涂阳肺结核患者的痰标本进行异烟肼和利福平耐药检测,以传统罗氏药敏试验为金标准,对基因芯片技术的检测效果进行评价.结果 在38例菌株中,用基因芯片法进行异烟肼耐药基因检测,与罗氏药敏的符合率为84.2％,对38例进行利福平耐药基因检测,符合率为89.5％.结论 基因芯片检测异烟肼和利福平的耐药性与罗氏药敏方法具有很好的一致性,具有简便快速,灵敏度高,特异性好的优点,对临床及时准确进行抗结核治疗具有重要意义.     

纳米金标记银染增强法快速检测痰液结核分枝杆菌

目的建立快速检测结核分枝杆菌(MTB)的纳米金标记银染增强法(金标记银染法)。方法以MTB的IS6110基因为靶基因,分别与互补的纳米金探针、生物素探针杂交,形成三明治杂交结构,然后借链霉亲合素将靶序列锚定在酶标孔内,通过纳米金催化的银染放大效应产生高灵敏的识别信号,在酶联仪上检测其吸光度(A)值。用本法、传统固体培养法、PCR法检测90例可疑患者痰标本并进行比对。结果金标记银染法检测限达1 pmol/L,90例临床标本的MTB检出率与PCR法均为42.2%(38/90);培养法MTB检出率为36.7%(33/90),金标记银染法与其检测结果差异无统计学意义(P0.05)。结论成功构建了金标记银染法,可直接应用于临床疑似肺结核患者痰标本的快速检测,具有简便快速、灵敏度高、特异性强等优点。     

联合检测ADA、LDH和CRP对结核性与恶性胸腔积液的诊断价值

目的探讨联合测定胸腔积液中的腺苷脱氨酶(ADA)、乳酸脱氢酶(LDH)和C反应蛋白(CRP)在结核性与恶性胸腔积液上鉴别诊断的价值。方法酶法和免疫比浊法检测59例结核性胸腔积液和17例恶性肿瘤积液中ADA、LDH和CRP的含量,并进行统计学分析。结果结核性胸腔积液中ADA和CRP水平显著高于恶性积液的值,具有统计学意义,P<0.01。但是,结核性胸腔积液中LDH含量低于恶性积液的LDH值,有显著性差异,P<0.01。结论胸腔积液中ADA、LDH和CRP的联合检测,对鉴别结核性与恶性胸腔积液有重要价值。     

Characterization of electrospun poly(L-lactide) and gold nanoparticle composite scaffolds for skeletal muscle tissue engineering

Traumatic injuries can interrupt muscle contraction by damaging the skeletal muscle and/or the peripheral nerves. The healing process results in scar tissue formation that impedes muscle function. Electrospinning and metal nanoparticles (Nps) can create a scaffold that will trigger muscle cell elongation, orientation, fusion, and striation. Poly(L-lactic acid) (PLLA) and gold (Au) Nps were electrospun to create three composite scaffolds, 7% Au-PLLA, 13% Au-PLLA and 21% Au-PLLA, and compared to PLLA alone. The scaffolds had a conductivity of 0.008 ± 0.003 S/cm for PLLA, 0.053 ± 0.015 S/cm for 7% Au-PLLA, 0.076 ± 0.004 S/cm for 13% Au-PLLA and 0.094 ± 0.037 S/cm for 21% Au-PLLA. Next, a cell study was conducted with rat primary muscle cells and all three Au-PLLA scaffolds. The first cell study showed low cell proliferation on all three of the Au-PLLA scaffolds; however, the second cell study showed that this was not due to Au Nps toxicity. Instead, low cell proliferation may be a marker for myotube differentiation and fusion. Values for the elastic modulus and yield stress for the Au-PLLA scaffolds on days 0, 7, 14, 21 and 28 were much higher than those for skeletal muscle tissue. Therefore, lower amounts of Au Nps may be utilized to create a biodegradable, biocompatible and conductive scaffold for skeletal muscle repair.     

心脏型脂肪酸结合蛋白定性检测对急性心肌梗死早期诊断的价值

目的:探讨心脏型脂肪酸结合蛋白(H-FABP)在急性心肌梗死(AMI)早期诊断中的价值。方法:运用胶体金免疫层析技术定性检测104例发病3h内胸痛患者的血H-FABP,比较其与心肌肌钙蛋白(cTnI)、肌酸激酶同功酶MB(CK-MB)、肌红蛋白(MYO)诊断早期AMI的敏感度、特异度。结果:入选患者中最终确诊为AMI患者50例,非AMI54例。H-FABP、cTnI、CK-MB和MYO检出AMI的敏感度分别为88.00%、68.00%、64.00%和84.00%,H-FABP显著高于cTnI和CK-MB(均P<0.05),与MYO比较无显著差异(P>0.05);检测特异度分别为90.74%、87.04%、85.19%和70.37%,H-FABP显著高于MYO(P<0.05),分别与cTnI和CK-MB比较均无显著差异(均P>0.05)。在四种心肌损伤标志物中,H-FABP的准确诊断指数最高。结论:H-FABP用于AMI早期诊断具有快速、简便,灵敏度和特异度高的特点,值得临床推广应用。     

建立用于检测H IV-1的基于核酸序列扩增的酶联免疫吸附测定法

目的：建立用于检测 HIV-1的基于核酸序列扩增的酶联免疫吸附测定（NASBA-ELISA）法。方法根据 HIV-1基因组长末端重复序列设计引物对 HIV-1 RNA进行基于核酸序列扩增（NASBA）,结合微孔板中液相杂交和酶标显色反应检测核酸扩增物,并对该检测方法的灵敏度、特异度及其对临床样本的检测结果进行评价。结果该方法可检测到0．1 fg/μL的RNA,且对 HIV-1具有特异性。在120例临床样本的检测中,其检测准确度高于常规的实时荧光定量反转录聚合酶链反应（fqRT-PCR）法。结论 NASBA-ELISA方法能灵敏并特异地检测 HIV-1。     

Targeting the mycobacterial envelope for tuberculosis drug development

The bacterium that causes tuberculosis, Mycobacterium tuberculosis, possesses a rather unique outer membrane composed largely of lipids that possess long-chain and branched fatty acids, called mycolic acids. These lipids form a permeability barrier that prevents entry of many environmental solutes, thereby making these bacteria acid-fast and able to survive extremely hostile surroundings. Antitubercular drugs must penetrate this layer to reach their target. This review highlights drug development efforts that have added to the slowly growing tuberculosis drug pipeline, identified new enzyme activities to target with drugs and increased the understanding of important biosynthetic pathways for mycobacterial outer membrane and cell wall core assembly. In addition, a portion of this review looks at discovery efforts aimed at weakening this barrier to decrease mycobacterial virulence, decrease fitness in the host or enhance the efficacy of the current drug repertoire by disrupting the permeability barrier.     

二步温控法聚合酶链反应检测脑脊液结核杆菌DNA

应用二步温控法聚合酶链反应（ＰＣＲ）检测脑脊液结核杆菌微量ＤＮＡ，并扩增出标准人型结核杆菌１５８ｂｐ的基因片段。实验结果表明：ＰＣＲ用于结核性脑膜炎患者脑脊液结核杆菌ＤＮＡ的检测，与传统检验方法相比较，具有敏感、快速、特异、高效等优点，是基因水平上检测结核杆菌的一项新技术，用于结核性脑膜炎的早期诊断，具有很强的实用价值，值得临床推广应用。     

不同核酸提取方法和新型冠状病毒核酸检测试剂室间质评结果分析

目的 分析2种不同核酸提取方法和2种新型冠状病毒核酸检测试剂检测室间质评样本的结果,进行性能比较.方法收集2020年江苏省临床检验中心和国家卫生健康委临床检验中心发放的新型冠状病毒核酸检测室间质量评价样本共15份,用2种常用核酸提取方法(磁珠法、核酸释放剂法)、2个厂家实时荧光定量PCR核酸扩增检测试剂检测,进行质评物...     

结核感染T细胞斑点试验在结核性疾病中的诊断价值

目的评价γ-干扰素释放分析结核感染T细胞斑点试验（T-SPOT,TB）检测在结核性疾病中的诊断价值。方法对仁济医院疑诊或待排结核患者进行T—SPOT．TB检测。结果T—SPOT．TB检测在结核性疾病阳性检出率为94．4％（17／18）,明显高于PPD的47．1％（8／17）、结核抗体检查的12．5％（2／16）、结核PCR的16．7％（2／12）、抗酸杆菌涂片的7．7％（1／13）、结核菌培养的27．3％（3／11）,差异有统计学意义（P〈0．05）。T-SPOT．TB检测诊断结核性疾病敏感性、特异性分别为94．4％、95．2％,显著优于PPD。结论T—SPOT．TB检测是诊断结核的快速敏感方法,在结核性疾病诊断中有重要价值。     

全血γ-干扰素释放试验与ELISA检测结核分枝杆菌抗体在痰菌阴性肺结核辅助诊断中的比较应用

目的：探讨γ-干扰素释放试验（interferon-gamma release assays, IGRA）与酶联免疫吸附试验（enzyme linked immunosorbent assay, ELISA）在痰菌阴性肺结核诊断中的应用价值。方法：收集93例痰菌阴性肺结核患者和40例健康体检者的肝素钠抗凝全血,采用IGRA试剂盒检测其血浆γ-干扰素含量,其血清标本应用ELISA试剂盒检测结核分枝杆菌抗体（Tuberculosis antibody, TBAb）,并将2种结果进行比较分析。将痰菌阴性肺结核患者分为老年组和中青年组进行比较,分析IGRA检测在不同年龄段间是否有差异。结果：IGRA诊断痰菌阴性肺结核,灵敏度为84.94%,特异度为90.00%,阳性预测值为95.18%,阴性预测值为72.00%;TBAb诊断痰菌阴性肺结核灵敏度为52.69%,特异度为85.00%,阳性预测值为67.20%,阴性预测值为40.96%。2种方法相比较,差异有统计学意义。不同年龄患者间,IGRA检测结果差异无统计学意义。结论：与ELISA相比,IGRA对诊断痰菌阴性肺结核有更高的灵敏度、特异度和阳性预测能力。     

含左旋氧氟沙星加母牛分支杆菌菌苗化疗方案治疗耐多药肺结核的护理

目的：探讨左氧氟沙星加母牛分支杆菌菌苗化疗方案治疗耐多药肺结核的疗效与护理。方法：将80例耐多药肺结核患者随机分为观察组和对照组各40例。观察组采用3DThZVAK＋M／15DThZV＋M方案,对照组采用3DThZEAK／15DThZE化疗方案。两组患者均同时配合支持治疗和对症护理。结果：疗程完成后,观察组的痰菌阴转率、病灶吸收率、空洞闭合率、症状改善率均优于对照组（P〈0．05）;药物不良反应发生率略低于对照组,但无统计学意义（P〉0．05）。结论：采用左氧氟沙星联合母牛分支杆菌菌苗治疗耐多药肺结核近期疗效确切,针对化疗药物不良反应给予及时有效的护理是取得治疗成功的重要保证。     

A comparative study of various methods for detection of IL28B rs12979860 in chronic hepatitis C

Interleukin-28B (IL28B) single-nucleotide polymorphisms (SNPs) constitute important host-related factors influencing the response rate to Hepatitis C virus (HCV) standard antiviral therapy. In the last few years, several new technologies for SNP detection have been developed. However, the sensitivity and specificity of various methods are different and needs evaluation. Five different methods (resolution melting curve [RMC], polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP], PCR-sequencing analysis, amplification refractory mutation system [ARMS], and zip nucleic acid probe-based real-time PCR [ZNA]) were developed for genotyping rs12979860 associated with IL28B. In this study, limit of detection (LD), costs and turnaround time of these methods were compared in 350 subjects. As for IL28B rs12979860 polymorphisms, 348/350 (99.4%) samples were consistent among the five methods, while results for 2/350 (0.57%) samples were concordant by ZNAs and PCR-sequencing, and discordant by other methods. Without considering the cost of DNA extraction, the price of each reaction for ARMS-PCR, RMC, PCR-RFLP, ZNA and PCR-sequencing were respectively: US$3.10, US$5.0, US$5.50, US$8.50 and US$17.0. RMC was the fastest method, while the ZNA method was easy to use, reliable and effective. Lower LD was determined to be 50–60 copies/μL for the PCR-RFLP, RMC and ARMS-PCR assays; whilst ZNA assay was able to detect 2–3 copies/μL. In conclusion, in the current study, all four methods are suitable for IL28B rs12979860 genotyping, but the ZNA assay can be a reliable tool. Due to its lower LD for SNP identification, this method is better than others for detecting this type of polymorphism.     

3项联合检测在急性冠状动脉综合征早期诊断中的应用价值

目的探讨肌酸激酶同工酶（CK—MB）、脂肪酸结合蛋白（H—FABP）联合肌钙蛋白（cTnT）检测在急性冠状动脉综合征（ACS）中的临床诊断价值。方法选取急性心肌梗死（AMI）患者58例和不稳定型心绞痛（UA）52例,同时选取同期行身体检查的健康体检者50例作为对照组,采用快速检验试弃1盒测定3组血清中H—FABP、cT—nT和CK—MB水平。结果AMI组与UA组相比,3种标记物水平显著上升,差异有统计学意义（P〈0．05）。H—FABP、cTnT和CKMB联合检测诊断ACS的准确性、灵敏度、特异性高于单一标记物及两种标记物联合检测,差异有统计学意义（P〈0．05）。结论H—FABP、cTnT、CK—MB联合检测能明显提高ACS诊断的准确性及灵敏度,具有较高的临床应用价值。     

Glucose-6 phosphate dehydrogenase deficiency: an easy and sensitive quantitative assay for the detection of female heterozygotes in red blood cells

Different methods for the quantitative determination of glucose-6-phosphate dehydrogenase (G-6-PD) were compared and one of them found to be highly precise. Maleimide inactivation of 6-phosphogluconate dehydrogenase (PGD) was investigated. It was shown that this inactivation is time-dependent and causes loss of assay precision. The most precise method was adapted to lysates of red blood cells from females, known to be heterozygote for G-6-PD deficiency and from non-deficient males and females. Heterozygote gene carriers were detected at a rate of 97.0%.     

Clinical evaluation of QuantiFERON®-TB Gold Plus directly compared with QuantiFERON®-TB Gold In-Tube and T-Spot®.TB for active pulmonary tuberculosis in the elderly

BackgroundReduced sensitivity of tuberculosis (TB) interferon-γ release assays (IGRAs) among the elderly has been reported, which is presumably due to diminished immune function. We evaluated the clinical performance of QuantiFERON®-TB Gold plus (QFT-Plus) compared with QuantiFERON®-TB Gold In-Tube (QFT-GIT) and T-Spot®.TB (T-SPOT) in the elderly.MethodsBlood samples for all three IGRAs were drawn at the same time from all the participants. Both CD4 and CD8 T-cell counts in patients’ peripheral blood were also measured.ResultsA total of 142 active pulmonary TB patients (median age: 84, interquartile range; 76–89 years) were recruited. The sensitivities of the tested IGRAs (excluding invalid/indeterminate cases) were as follows: QFT-Plus, 93.6%; QFT-GIT, 91.4%; and T-SPOT 68.1%. QFT-Plus displayed significantly higher sensitivity than T-SPOT (p < 0.00001). All three IGRAs exhibited the same specificity (100%), as assessed using blood samples from healthy, low TB-risk individuals (n = 118; median age: 39, IQR; 32–47 years). Positivity in 43 active TB patients with CD4 T-cell counts <200/μL, 39 of whom were ≥80 years of age, was as follows: QFT-Plus, 83.7%; QFT-GIT, 74.4%; and T-SPOT, 58.1%. The difference between TB2-TB1 of the QFT-Plus assay was statistically correlated with CD8 but not CD4 T-cell counts in blood (r = 0.193, p = 0.0298).ConclusionsQFT-Plus showed high performance in the detection of TB infection in patients irrespective of their advanced age (≥80 years) or lower CD4 counts. QFT-Plus can be useful for the diagnosis of TB infection in all patients, including those who are elderly and/or immunocompromised.     

“Nanoantibiotics”: A new paradigm for treating infectious diseases using nanomaterials in the antibiotics resistant era

Despite the fact that we live in an era of advanced and innovative technologies for elucidating underlying mechanisms of diseases and molecularly designing new drugs, infectious diseases continue to be one of the greatest health challenges worldwide. The main drawbacks for conventional antimicrobial agents are the development of multiple drug resistance and adverse side effects. Drug resistance enforces high dose administration of antibiotics, often generating intolerable toxicity, development of new antibiotics, and requests for significant economic, labor, and time investments. Recently, nontraditional antibiotic agents have been of tremendous interest in overcoming resistance that is developed by several pathogenic microorganisms against most of the commonly used antibiotics. Especially, several classes of antimicrobial nanoparticles (NPs) and nanosized carriers for antibiotics delivery have proven their effectiveness for treating infectious diseases, including antibiotics resistant ones, in vitro as well as in animal models. This review summarizes emerging efforts in combating against infectious diseases, particularly using antimicrobial NPs and antibiotics delivery systems as new tools to tackle the current challenges in treating infectious diseases.