共查询到17条相似文献,搜索用时 93 毫秒
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目的:建立一种快速检测新型隐球菌的方法。方法根据新型隐球菌的18 S rRNA基因序列设计内外引物各一对,在不同温度下对新型隐球菌进行环介导等温扩增,以探明环介导等温扩增法检测新型隐球菌最适宜的温度。对新型隐球菌、其他几种常见引起脑膜炎的病原菌以及部分种类的念珠菌进行扩增,以研究环介导等温扩增技术检测隐球菌的特异性。将新鲜培养的新型隐球菌稀释后加入非隐球菌感染患者的脑脊液中,模拟脑膜炎感染后的脑脊液环境,以研究环介导等温扩增技术检测新型隐球菌的敏感性。结果环介导等温扩增法检测新型隐球菌在60℃~65℃条件下均能实现良好扩增。环介导等温扩增法对其他病原菌的检测结果均为阴性,仅新型隐球菌的检测结果为阳性,特异性达到100%。用环介导等温扩增法从脑脊液中检测新型隐球菌的最低检出限为102 CFU/ml。用环介导等温扩增技术检测新型隐球菌性脑膜炎,能节省培养及传统鉴定的时间,从DNA的提取至反应结束在2~3h内即可完成,大大减少了诊断所需时间。结论环介导等温扩增是一种灵敏度高,特异性强,耗时短,且操作简便的检测新型隐球菌感染的方法。 相似文献
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前列腺癌是男性泌尿系统常发且严重威胁男性健康的恶性肿瘤,且预后较差。由于其临床上缺乏明显的症状且影像学也缺乏特殊表现,等到发现时多已进展到晚期,唯一能避免此结果的方法就是通过有效、准确的检测从而早期诊断,并进行相应治疗。目前,关于以DD3作为前列腺癌的肿瘤标记物进行检测是为研究热点,研究者对样本的选取以及相关检测方法均进行了探索,本文将对这一最新研究探索进行综述。 相似文献
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DD3 mRNA在前列腺癌组织中定量表达分析 总被引:8,自引:0,他引:8
目的:探讨DD3基因在前列腺组织中表达的临床意义。方法:用荧光定量RT-PCR方法对21例前列腺癌(PCa)组织、27例非前列腺部位的肿瘤组织、39例良性前列腺增生(BPH)组织和15例正常前列腺组织中的DD3的表达进行了定量分析,用ROC曲线对DD3 mRNA诊断PCa的性能进行了分析。结果:27例非前列腺组织中均未检测到DD3基因的表达。DD3在PCa组、BPH组和正常前列腺组表达量的中位数分别为7.2×106、2.5×104、1.5×104拷贝/mg组织。PCa组较BPH组和正常前列腺组DD3的表达量显著增高(P<0.01),而BPH组和正常前列腺组间则差异无显著性(P>0.05)。DD3表达量与临床分期和分化程度之间均无明显相关性。DD3 mRNA曲线下面积(AUC-ROC)为0.937(95%CI:0.879~0.995)。当临界值为1.4×105拷贝/mg组织时,灵敏度、特异度、准确度、阳性预测值(PPV)、阴性预测值(NPV)、阳性拟然比(+LR)、阴性拟然比(-LR)分别为90.5%、85.0%、86.7%、76.0%、94.3%、6.03和0.11。结论:DD3 mRNA的表达仅限于前列腺组织,具有良好的组织特异性。DD3 mRNA表达在PCa组织中显著升高,在正常前列腺和BPH组织中差异无显著性。DD3 mRNA可作为PCa诊断的良好标志物,在PCa早期诊断、微转移诊断、预后判断、指导治疗等方面也具有潜在的应用价值。 相似文献
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前列腺癌患者尿液中DD3 mRNA检测及临床意义 总被引:1,自引:0,他引:1
目的:观察PCa患者尿液中DD3 mRNA的表达,并讨论其临床意义。方法:从48例PCa患者、23例BPH患者和9例健康男性志愿者(对照组)前列腺按摩后初始尿液中分离细胞,采用RT-PCR方法检测其中DD3mRNA的表达情况。结果:BPH和对照组未见DD3 mRNA阳性表达,而PCa患者DD3 mRNA阳性率为81.3%(39/48),两者差异具有统计学意义(P<0.01)。结论:尿液中DD3 mRNA的检测有望成为早期诊断PCa的一种无创、简单、敏感的方法,也有望成为PCa治疗后监测的一种方法。 相似文献
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目的检测前列腺癌(PCa)患者前列腺液中DD3基因的表达,讨论其临床意义。方法收集31例PCa患者、59例前列腺增生(BPH)患者前列腺液,离心取细胞沉淀物,提取总RNA,采用逆转录-聚合酶链反应(RT-PCR)琼脂糖电泳法方法检测其中DD3 mRNA的水平。同时用2χ检验分析比较不同Gleason分级的PCa患者DD3 mRNA阳性率是否存在差异。结果 PCa患者前列腺液中DD3 mRNA阳性率(77.42%)明显高于BPH患者(5.08%),两者具有显著性差异(P〈0.01)。DD3 mRNA表达阳性率诊断PCa的灵敏度为77.42%,特异度为94.91%,不同Gleason分级的PCa患者DD3 mRNA表达阳性率无显著统计学差异(P〉0.05)。结论检测前列腺液中DD3 mRNA诊断PCa是一种有效的无侵入性的良好方法。 相似文献
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目的:探讨PSA值增高患者前列腺按摩后尿液沉渣中DD3mRNA检测在前列腺癌诊断中的应用价值.方法:在前列腺穿刺活检前或术前麻醉后收集患者前列腺按摩后患者的尿液,离心取细胞沉淀物,用逆转录-聚合酶链反应(RT-PCR)法和荧光实时定量逆转录-聚合酶链反应(Real time RT-PCR)方法检测DD3mRNA和PSAmRNA含量,以PSAmRNA作为管家基因校正沉渣中的前列腺细胞,以DD3mRNA/PSAmRNA比值表示DD3mRNA含量.SPSS13.0分析相关数据,以穿刺活检或术后病理检查结果为金标准,同时探讨了前列腺按摩后尿液中DD3Mrna含量与前列腺癌病理分级之间的关系.结果:PSA值增高患者中前列腺癌患者前列腺液DD3mRNA表达量明显高于BPH,差异具有统计学意义.不同病理分级之间DD3mRNA的含量差异无统计学意义(P>0.05).结论:PSA值增高的患者行前列腺按摩后尿液沉渣中DD3mRNA含量检测,较前列腺活检的损伤更小,作为前列腺癌的一种非损伤性诊断方法具有良好的应用前景. 相似文献
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目的评价外周血前列腺癌抗原基因-3(prostate cancer antigen3,PCA3 mRNA)和前列腺特异性抗原基因(prostate specific antigen,PSA mRNA)联合检测对前列腺癌(PCa)早期诊断的价值,及决定是否进行前列腺穿刺活检。方法纳入41例PCa和69例前列腺增生(BPH)患者的外周血,采用RT—PCR方法对其PCA3mRNA和PSAmRNA进行检测,通过受试者工作特征(ROC)曲线评价其在预测前列腺穿刺活检结果和PCa早期诊断的价值。结果PCa组外周血PCA3mRNA含量明显高于BPH组[2362(〈30~7421)拷贝/ml比〈35拷贝/ml,z=-6.66,P〈0.01],而PSAmRNA含量也明显高于BPH组[3425(908~36639)拷贝/ml比〈220拷贝/ml,z=-6.40,P〈0.01];ROC曲线显示当PCA3mRNA和PSAmRNA临界值分别为846拷贝/ml和280拷贝/ml时,诊断早期PCa结果为73.2%(30/41)、85.4%(35/41),特异度分别为87.0%(60/69)、78.3%(54/69);而联合检测时其敏感度可增至90.2%(37/41),特异度上升为89.9%(62/69)。结论外周血PCA3mRNA和PSAmRNA检测都是PCa诊断的良好指标,而它们的联合检测可弥补PCA3mRNA敏感性低和PSAmRNA特异性低的缺点,更有利于早期PCa诊断。 相似文献
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目的探讨DD3 mRNA和PSA mRNA在前列腺组织中表达的临床意义及诊断前列腺癌(PCa)的价值。方法荧光定量RT—PCR法分析21例PCa组织、39例良性前列腺增生(BPH)组织DD3 mRNA和PSA mRNA的表达,ROC曲线对DD3 mRNA、PSA mRNA和DD3 mRNA/PSA mRNA比值诊断PCa的价值进行分析。结果PCa组织中DD3 mRNA、PSA mRNA表达量和DD3 mRNA/PSA mRNA比值均显著高于BPH组织,差异有统计学意义(P<0.01)。不同临床分期和分化程度之间DD3 mRNA、PSA mRNA和DD3 mRNA/PSA mRNA比值差异无统计学意义(P值均>0.05)。ROC曲线分析结果显示,DD3 mRNA、PSA mRNA和DD3 mRNA/PSA mRNA的曲线下面积分别为0.937、0.755和0.839。当DD3 mRNA、PSA mRNA和DD3 mRNA/PSA mRNA临界值分别为1.4×105拷贝/mg组织、3.0×107拷贝/mg组织和5.0×10-3时,敏感性分别为90.5%、81.0%和81.0%,特异性分别为85.0%、62.0%和66.7%。若将DD3 mRNA和PSA mRNA联合用于PCa的诊断,其特异性与DD3 mRNA相同,为85.0%,敏感性可达100.0%。结论PCa组织中DD3 mRNA和PSA mRNA表达量均增加,但组织中DD3 mRNA的定量检测更具诊断价值,联合检测有利于提高诊断敏感性,对PCa的诊断具有一定临床应用价值。 相似文献
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环氧化酶2 mRNA在前列腺癌组织中的表达 总被引:4,自引:4,他引:4
目的:探讨环氧化酶2(COX-2)mRNA在前列腺癌(PCa)组织中的表达及其意义。方法:采用RT-PCR方法检测32例PCa组织中COX-2 mRNA的表达并测定COX-2/GAPDH的比值,7例正常前列腺组织作为对照。结果:7例正常前列腺组织无COX-2 mRNA表达。COX-2/GAPDH与PCa G leason评分、分期呈正相关(P均<0.05)。结论:COX-2在PCa的发生发展中起重要的正性调节因子的作用。COX-2与PCa的恶性程度和进展有关,可作为判断PCa预后的指标。 相似文献
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目的:提取并鉴定已构建的EB病毒表达载体pDR2-TK,利用脂质体介导的基因转染技术将pDR2-TK转染人前列腺癌细胞并对单纯疱疹胸苷激酶(HSV-TK)表达状况进行检测。方法:采用DNA大量制备及纯化系统提取pDR2-TK,酶切和DNA测序进行鉴定,采用阳离子脂质体法将pDR2-TK导入激素非依赖性人前列腺癌细胞系PC-3m,逆转录PCR(RT-PCR)法和SABC免疫组化法检测TK mRNA和蛋白的表达。结果:扩增提取的质粒经PstⅠ和EcoR Ⅴ酶切后各获得4个及2个片段,与原基因酶切图谱一致;所提取质粒PCR产物经DNA测序,与NCBI公布的HSV-TK基因序列对照,证实所提取质粒含目的基因序,旨质体法转染PC-3m细胞后,mRNA和蛋白均有HSV-TK的表达,其蛋白表达率约为22%。结论:pDR2-TK质粒含有目的的基因HSV-TK,阳离子脂质体法可将pDR2-TK导入人前列腺癌细胞并获得较高效率的表达。 相似文献
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J. Yang G. Guan Q. Niu Z. Liu Y. Li J. Liu M. Ma Q. Ren A. Liu J. Luo H. Yin 《Transboundary and Emerging Diseases》2013,60(3):238-244
A loop‐mediated isothermal amplification (LAMP) assay was developed to detect Borrelia burgdorferi s. l. in ticks, which is a pathogen that causes Lyme disease. Cross‐reactions with Chlamydia psittaci, Mycoplasma mycoides subsp. capri and some tick‐borne pathogens were excluded. Analytical sensitivity of LAMP showed its detection limit was from 0.02 to 0.2 pg of DNA in detection of the reference samples at 65°C for 40 min. The performance of LAMP was assessed by testing 110 samples from susceptible tick species and comparing the results with conventional and nested PCR tests previously described. The results demonstrated that LAMP was significantly more sensitive than the conventional PCR (32.7% versus 15.5%, P < 0.05) and slightly more sensitive, although not significantly so, than nested PCR (32.7% versus 26.4%, P > 0.05). The assay was used to analyse a total of 1052 ticks collected from eight provinces in China. The results showed that the infection rates of B. burgdorferi s. l. varied from 12.5% to 88.9% across the different geographical sites. Selected positive samples were subjected to sequencing and sequence analysis for conformation of the accuracy of the assay. Here we report a highly sensitive, specific and easy diagnostic assay based on LAMP technology. These data indicate that LAMP is a useful approach for detecting B. burgdorferi s. l. in field‐collected ticks and has the potential as an alternative tool for the ecological and epidemiological surveillance of Lyme disease. 相似文献
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《Transboundary and Emerging Diseases》2018,65(3):597-601
In this study, a rapid and specific assay for the detection of porcine circovirus type 3 (PCV 3) was established using loop‐mediated isothermal amplification (LAMP ). Four primers were specifically designed to amplify PCV 3. The LAMP assay was effectively optimized to amplify PCV 3 by water bath at 60°C for 60 min. The detection limit was approximately 1 × 101 copy in this LAMP assay. Compared to porcine circovirus type 2 (PCV 2), both gE and gD genes of pseudorabies virus (PRV ) and porcine parvovirus (PPV ), the LAMP assay showed a high specific detection of PCV 3. A visible detection method was developed using SYBR Green I to recognize the results rapidly. Based on the detection of 20 clinical tissue samples, the LAMP assay was more practical and convenient than classical PCR due to its simplicity, high sensitivity, rapidity, specificity, visibility and cost efficiency. 相似文献
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Hiroshi Maeda Yoichi Arai Kazutoshi Okubo Yoshitaka Aoki Takashi Okada Shinya Maekawa 《International journal of urology》1998,5(4):343-348
Background : This study evaluated the free to total serum prostate specific antigen (f/t PSA) ratio and prostate specific antigen density (PSAD) in detecting prostate cancer in Japanese males with a PSA level between 2.5 and 20.0 ng/mL in a community-based urology practice.
Methods : Twenty-six patients with clinically localized prostate cancer and 44 patients with histologically-proven benign prostatic hyperplasia (BPH) were studied. The serum levels of free PSA (fPSA) and total (t) PSA were determined using a chemiluminescent enzyme immunoassay. The f/t PSA ratio was calculated by dividing the fPSA value by the total PSA value and was compared with the PSA and PSAD via the receiver operating characteristic (ROC) curves.
Results: Patients with prostate cancer had a significantly lower f/t PSA ratio than patients with BPH. The PSAD was a superior diagnostic tool over PSA (P < 0.01) when analyzed by ROC curves. The f/t PSA ratio was also superior to PSA, but lacked significance (P=0.12), and similarly, the PSAD was superior, but not significant, to the f/t PSA ratio. Using a cut-off value of 0.1 9, the PSAD had a sensitivity of 81% and a specificity of 82%. With a cut-off value of 14.0%, the f/t PSA ratio had a sensitivity of 81% and a specificity of 66%.
Conclusion: This study showed that PSAD alone improved cancer detection significantly better than PSA. However, it is still unclear whether the f/t PSA ratio is superior to PSA or PSAD in the discrimination between BPH and prostate cancer in Japanese male patients. 相似文献
Methods : Twenty-six patients with clinically localized prostate cancer and 44 patients with histologically-proven benign prostatic hyperplasia (BPH) were studied. The serum levels of free PSA (fPSA) and total (t) PSA were determined using a chemiluminescent enzyme immunoassay. The f/t PSA ratio was calculated by dividing the fPSA value by the total PSA value and was compared with the PSA and PSAD via the receiver operating characteristic (ROC) curves.
Results: Patients with prostate cancer had a significantly lower f/t PSA ratio than patients with BPH. The PSAD was a superior diagnostic tool over PSA (P < 0.01) when analyzed by ROC curves. The f/t PSA ratio was also superior to PSA, but lacked significance (P=0.12), and similarly, the PSAD was superior, but not significant, to the f/t PSA ratio. Using a cut-off value of 0.1 9, the PSAD had a sensitivity of 81% and a specificity of 82%. With a cut-off value of 14.0%, the f/t PSA ratio had a sensitivity of 81% and a specificity of 66%.
Conclusion: This study showed that PSAD alone improved cancer detection significantly better than PSA. However, it is still unclear whether the f/t PSA ratio is superior to PSA or PSAD in the discrimination between BPH and prostate cancer in Japanese male patients. 相似文献
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经会阴B型超声引导下前列腺穿刺活检的临床价值研究 总被引:1,自引:0,他引:1
目的:探讨经会阴扇型B型超声引导下前列腺6针穿刺活检术诊断前列腺癌的临床价值。方法:对经直肠指检或经腹部B超检查发现前列腺结节、血清总前列腺特异性抗原(tPSA)在4μg/L以上或游离PSA(fPSA)/tP-SA<0.16的可疑前列腺癌104例患者,经会阴扇型B超引导下18G自动穿刺活检针行双侧叶6点法穿刺,对穿刺的阳性率和并发症及影响穿刺阳性率的因素进行分析。结果:经病理诊断,检出前列腺癌24例,检出率23%,前列腺癌分级评分中位数为7分,高分化癌(2~4分)、中分化癌(5~7分)和低分化癌(8~10分)分别为12.5%(3/24)、62.5%(15/24)和25%(6/24);其余80例为良性前列腺增生(BPH)。术后短暂和轻度的肉眼血尿5例(4.8%),均在1~3 d后缓解,4例(3.8%)发热37.2℃~38.0℃,术后会阴部轻度不适5例(4.8%)。术后无1例出现血便、血精、前列腺脓肿、高热、败血症、急性尿潴留等严重并发症。经分析发现tPSA、fPSA、fPSA/tPSA、前列腺抗原密度(PS-AD)和前列腺体积是影响前列腺穿刺阳性率的重要因素(P<0.05),经会阴穿刺优势主要反映在tPSA≥10μg/L、fP-SA≥2μg/L、fPSA/tPSA<0.16、PSAD≥0.2和前列腺体积<40 m l时提示应行会阴穿刺术。结论:经会阴扇型B型超声引导下6针前列腺穿刺活检,是一种安全准确的前列腺癌检出方法。 相似文献
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经直肠超声引导穿刺活检诊断早期前列腺癌漏诊原因的分析 总被引:1,自引:1,他引:0
目的:分析经直肠超声(TRUS)引导下穿刺活检诊断前列腺癌的漏诊原因,减少漏诊率,提高诊断率。方法:80例疑似前列腺癌的良性前列腺增生(BPH)患者行TRUS引导下穿刺活检,结果均为阴性,均行前列腺电切术(TURP),术后标本行病理检查。结果:25例术后病理报告为前列腺癌,漏诊率31.25%(25/80)。其中10例行经会阴前列腺癌根治术、8例行手术去势、7例行药物去势。结论:TRUS引导穿刺活检诊断前列腺癌存在一定的漏诊,多次或多点穿刺活检可以减少漏诊率。 相似文献
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目的:观察葡萄籽提取物(GSE)对前列腺癌PC-3细胞的生长抑制作用。方法:经预试选用不同浓度(100、200、300μg/ml)的GSE分别作用PC-3细胞24、48、72h,以原代培养的1~3日龄SD大鼠肾细胞作为正常对照;采用四唑氮蓝(MTT)显色法检测GSE对PC-3细胞和SD大鼠肾细胞的生长抑制作用。结果:GSE以浓度和时间依赖性方式抑制PC-3细胞生长(P<0.01),对原代培养SD大鼠肾细胞生长仅有轻度抑制作用。结论:GSE可抑制前列腺癌PC-3细胞生长,有可能成为治疗前列腺癌的一种新药物。 相似文献