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首页> 外文期刊>Journal of Clinical Microbiology >New Commercially Available PCR and Microplate Hybridization Assay for Detection and Differentiation of Human Polyomaviruses JC and BK in Cerebrospinal Fluid, Serum, and Urine Samples
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New Commercially Available PCR and Microplate Hybridization Assay for Detection and Differentiation of Human Polyomaviruses JC and BK in Cerebrospinal Fluid, Serum, and Urine Samples

机译：用于检测和区分脑脊液，血清和尿液样本中的人类多瘤病毒JC和BK的新型商用PCR和微孔板杂交测定

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JC and BK human polyomaviruses (family Polyomaviridae) may cause severe neurological or urinary tract pathologies in immunocompromised hosts. In the present study, we evaluated a new commercially available PCR and microplate colorimetric hybridization assay for the standardized differential detection of JC virus (JCV) and BK virus (BKV) genomes in clinical samples. This JC/BK Consensus test was first evaluated by testing serial dilutions of JCV or BKV plasmid DNA standards and was then compared with an in-house reference PCR assay for the detection of JC and BK virus genomes in 70 cerebrospinal fluid (CSF) samples of patients with neurological disorders and in 75 serum or plasma samples and 125 urine samples of renal graft recipients. This new test allowed a limit of detection of 10 copies and 1 copy of JC and BK virus genomes, respectively, and was able to differentiate various levels of JCV, BKV, and mixed JCV and BKV DNA genomes in a single reaction tube. Our results showed 100% specificity and sensitivity for the JC/BK Consensus test with CSF samples. With serum or plasma samples, this test had a sensitivity and a specificity of 100% for both JCV and mixed JCV and BKV DNA detection and a sensitivity and a specificity of 100 and 97.8% for BKV DNA detection, respectively. With urine samples, the sensitivity and specificity were 100 and 96.6%, respectively, for JCV DNA detection; 100 and 89.4%, respectively, for BKV DNA detection; and 44.4 and 100%, respectively, for mixed JCV and BKV DNA detection. In conclusion, our data indicate that this new test, the JC/BK Consensus test, is valuable for the sensitive and specific differential detection of single JCV and BKV infections in CSF, serum or plasma, and urine samples. The use of this reliable PCR assay would improve the routine virological diagnosis as well as the clinical care of immunocompromised patients with polyomavirus-related pathologies.
机译：JC和BK人类多瘤病毒（oly> Polyomaviridae 家族）可能在免疫受损的宿主中引起严重的神经系统或泌尿道疾病。在本研究中，我们评估了一种新的市售PCR和微孔板比色杂交测定法，用于临床样品中JC病毒（JCV）和BK病毒（BKV）基因组的标准化差异检测。这项JC / BK共识测试首先通过测试JCV或BKV质粒DNA标准品的系列稀释液进行评估，然后与内部参考PCR分析法进行比较，以检测70例脑脊液（CSF）样本中的JC和BK病毒基因组。有神经系统疾病的患者，以及肾移植受者的75份血清或血浆样本和125份尿样。这项新测试允许分别检测10个拷贝和1个拷贝的JC和BK病毒基因组，并且能够在单个反应管中区分不同水平的JCV，BKV以及混合的JCV和BKV DNA基因组。我们的结果表明，使用CSF样品进行JC / BK共识测试的特异性和敏感性为100％。对于血清或血浆样品，该测试对JCV以及JCV和BKV混合DNA检测的灵敏度和特异性分别为100％，对BKV DNA检测的灵敏度和特异性分别为100和97.8％。尿液样本对JCV DNA检测的敏感性和特异性分别为100和96.6％。 BKV DNA检测分别为100％和89.4％; JCV和BKV DNA混合检测分别为44.4和100％。总之，我们的数据表明，这种新的检测方法，即“ JC / BK共识性检测”，对于灵敏和特异的差异检测在脑脊液，血清或血浆以及尿液样本中的单个JCV和BKV感染非常有价值。使用这种可靠的PCR检测方法将改善常规病毒学诊断以及患有多瘤病毒相关病理的免疫功能低下患者的临床护理。

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