Morshed Kamal, Polz-Dacewicz Małgorzata, Szymański Marcin, Smoleń Agata
Department of Otolaryngology Head and Neck Surgery, University of Lublin, Lublin, Poland.
Folia Histochem Cytobiol. 2010 Sep 30;48(3):398-402. doi: 10.2478/v10042-010-0025-z.
The use of formalin-fixed paraffin-embedded (FFPE) tissues for HPV DNA detection by PCR from biopsy materials is not entirely clear in retrospective studies. The aim of our study was to evaluate the usefulness and efficiency of FFPE tissues from laryngeal cancer (LSCC) in HPV detection by immunohistochemistry reaction (IHC) and PCR-DNA enzyme immunoassay method (PCR/DEIA) and to compare with HPV detection from DFT. HPV-DNA was amplified from 54 FFPE tissues from LSCC specimens by the short PCR fragment (SPF10) primer set using PCR/DNA method and monoclonal anti Human Papillomavirus antibodies in IHC. In the same patients 54 specimens were collected and immediately deep-frozen and stored at (-70°C) to (-80°C). All the FFPE and deep-frozen tissue (DFT) specimens were positive for β-globin amplification. HPV was detected by two methods (SPF10 PCR/DEIA and IHC) in 14 (25.92%) out of 54 specimens from FFPE. Significant differences were found between the HPV detection using PCR/DEIA method and IHC method in FFPE tissues. The comparative analysis of the 54 samples after assuming PCR method in FFPE tissues showed accuracy of 92.6%, sensitivity of 90.5% and specificity of 93.9%. The FFPE tissues method has high sensitivity, specificity and accuracy when used to detect HPV DNA by PCR reaction and it is comparable to DFT results. DNA quality of FFPE samples is adequate and it can be used in HPV-DNA detection and in retrospective studies on LSCC.
在回顾性研究中，通过聚合酶链反应（PCR）从活检材料中使用福尔马林固定石蜡包埋（FFPE）组织检测人乳头瘤病毒（HPV）DNA的情况尚不完全清楚。我们研究的目的是评估来自喉癌（LSCC）的FFPE组织在通过免疫组织化学反应（IHC）和PCR-DNA酶免疫测定法（PCR/DEIA）检测HPV中的实用性和效率，并与从深冻组织（DFT）中检测HPV进行比较。使用PCR/DNA方法的短PCR片段（SPF10）引物组和IHC中的单克隆抗人乳头瘤病毒抗体，从54个LSCC标本的FFPE组织中扩增HPV-DNA。在同一患者中，收集了54个标本并立即深度冷冻，保存在（-70°C）至（-80°C）。所有FFPE和深冻组织（DFT）标本的β-珠蛋白扩增均为阳性。通过两种方法（SPF10 PCR/DEIA和IHC）在54个FFPE标本中的14个（25.92%）中检测到HPV。在FFPE组织中，使用PCR/DEIA方法和IHC方法检测HPV之间存在显著差异。在假设FFPE组织采用PCR方法后对54个样本进行的比较分析显示，准确率为92.6%，灵敏度为90.5%，特异性为93.9%。FFPE组织方法在通过PCR反应检测HPV DNA时具有高灵敏度、特异性和准确性，并且与DFT结果相当。FFPE样本的DNA质量足够，可用于HPV-DNA检测和LSCC的回顾性研究。
