三种多重实时PCR检测单纯疱疹病毒1型和2型DNA的比较

Anna Majewska, Maciej Przybylski, Paulina Machura, Graźyna Mlynarczyk, Tomasz Dzieciqtkowski
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引用次数: 0

摘要

简介:1型和2型单纯疱疹病毒(HSV-1和HSV-2)在世界范围内引起广泛感染,病程和强度不同。虽然这种病毒引起的疾病主要涉及健康的儿童和成人,但HSV感染对免疫缺陷的人来说要危险得多。本工作的目的是比较两种qPCR检测HSV-1/2 DNA的诊断价值,基于TaqMan*和HybProbes化学与商业HSV-1/2 Qual Kit。方法:在LightCycler 2.0热循环仪上检测51份临床样本的DNA是否存在HSV-1/2序列,采用两种内部开发的多重实时PCR检测方法和SCORPIONSTM引物进行商业检测。结果:所采用的分子生物学检测方法均具有较高的特异性和敏感性。统计学上,TaqMan*和HybProbes的化学方法与基于SCORPIONSTM的商业方法的实时PCR检测灵敏度无显著差异(P>0.05)。结论:所有检测方法特异性高,可用于临床样品中HSV-1/2 DNA的同时检测和鉴别。qPCR检测的高检出率和短持续时间对免疫功能低下患者具有重要意义,需要快速应用有效安全的治疗方法。在非定形感染和出现非特异性和全身性症状时也很重要。
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[Comparison of three multiplex real-time PCR assays for the detection of herpes simplex viruses type 1 and type 2 DNA].

Introduction: Herpes simplex viruses type 1 and type 2 (HSV-1 and HSV-2) cause widespread infection worldwide with different course and intensity. Although the disease caused by this viruses mainly concern healthy children and adults, the HSV infections are much more dangerous for people with immunodeficiencies. The aim of this work was to compare the diagnostic value of two qPCR methods for detection HSV-1/2 DNA, based on TaqMan* and HybProbes chemistry with commercial HSV-1/2 Qual Kit.

Methods: DNA from 51 clinical samples was tested for presence of HSV-1/2 sequences on LightCycler 2.0 thermocycler, using two ,,in-house" developed multiplex real-time PCR assays and commercial test using SCORPIONSTM primers.

Results: The results showed high specificity and sensitivity of all molecular biology tests used. Statistically, there was no significant difference in the sensitivity of real-time PCR assays when using TaqMan* and HybProbes' chemistry and when using the commercial SCORPIONSTM based method (P>0.05).

Conclusions: Obtained results show that all tested methods are highly specific and can possibly be used to simultaneously detect and differentiate HSV-1/2 DNA in clinical samples. The high detection rate and short duration of qPCR assayas has great importance for immunocompromised patients where quick application of effective and safe treatment is necessary. It is also important in the event of amorphous form of the infection and the occurrence of nonspecific and generalized symptoms.

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