Midak-Siewirska Anna, Karabin Karolina, Chudzik Emilia, Dzieciatkowski Tomasz, Przybylski Maciej, Majewska Anna, Łuczak Mirosław, Młynarczyk Grazyna
Katedra i Zakład Mikrobiologii Lekarskiej, Warszawski Uniwersytet Medyczny.
Med Dosw Mikrobiol. 2010;62(1):85-92.
Herpes simplex virus type 1 is a member of the Alphaherpesviridae subfamily, as it can infect both skin and nerves and develop latent infection within the dorsal root and trigeminal ganglia. Infection with this virus is common and causes a wide range of clinical syndromes. Although HSV-1 infect healthy children and adults, disease is more severe and extensive in the immunocompromised individuals. The goal of the study was development of real-time PCR assay for detection of herpes simplex virus type 1 DNA in clinical samples, using primers targeting a conserved region of the viral DNA glycoprotein G gene and a specific TaqMan hydrolyzing probe. The analytical sensitivity of assay was tested using serial dilutions of HSV-1 DNA in range between 10(0) and 10(-6) (4,35 x 10(5) - 4,00 x 10(1) copies/ml). Fifteen cell line isolates and twenty plasma samples taken from a group of adult recipients of allogeneic HSCT were tested for the presence of HSV-1 DNA in the LightCycler system. For comparison commercial quantitative HSV-1/2 LC PCR Kit (Artus/Qiagen) was used, according to the manufacturer's instructions. Both LightCycler assays, including in-house real-time PCR, detected HSV-1 DNA in 23 specimens. The conclusion is that developed TaqMan-based probe real-time PCR test is very reliable and valuable for detection of HSV-1 viremia in different kind of samples. The high level of sensitivity and accuracy provided by the LightCycler instrument is favorable for the use of this method in the detection of herpes simplex virus 1 DNA also in clinical specimens.
单纯疱疹病毒1型是α疱疹病毒亚科的成员，因为它既能感染皮肤和神经，又能在背根神经节和三叉神经节内形成潜伏感染。这种病毒感染很常见，会引发多种临床综合征。虽然HSV-1可感染健康儿童和成人，但在免疫功能低下的个体中，疾病会更严重且范围更广。该研究的目的是开发一种实时PCR检测方法，用于检测临床样本中的单纯疱疹病毒1型DNA，使用针对病毒DNA糖蛋白G基因保守区域的引物和一种特异性TaqMan水解探针。使用10(0)至10(-6)（4.35×10(5) - 4.00×10(1)拷贝/毫升）范围内的HSV-1 DNA系列稀释液测试该检测方法的分析灵敏度。对15株细胞系分离株和从一组异基因造血干细胞移植成年受者采集的20份血浆样本进行检测，以确定在LightCycler系统中是否存在HSV-1 DNA。为作比较，按照制造商说明使用商业定量HSV-1/2 LC PCR试剂盒（Artus/Qiagen）。两种LightCycler检测方法，包括内部实时PCR检测，均在23份标本中检测到HSV-1 DNA。结论是，所开发的基于TaqMan探针的实时PCR检测方法对于检测不同类型样本中的HSV-1病毒血症非常可靠且有价值。LightCycler仪器提供的高灵敏度和准确性有利于该方法在临床标本中检测单纯疱疹病毒1型DNA时的应用。
