Hantz S, Goudard M, Marczuk V, Renaudie J, Dussartre C, Bakeland D, Denis F, Alain S
Laboratoire de bactériologie-virologie-hygiène, CHU de Limoges, 2, avenue Martin-Luther-King, 87042 Limoges, France.
Pathol Biol (Paris). 2010 Apr;58(2):179-83. doi: 10.1016/j.patbio.2009.07.019. Epub 2009 Oct 27.
The objective of this study is to validate the use of test INNO-LIPA HPV Genotyping Extra (Innogenetics) on liquid cytology media EasyFix Labonord by comparing the extraction kit QIAamp DNA Blood Mini Kit (Qiagen) and an automated method, Nuclisens easyMAG (Biomérieux).
Thirty-two samples were typed by the technique Hybrid Capture 2 (HC2) Digene (Qiagen). DNA was extracted through manual or automated extraction and quality controlled PCR "HLA". Typing was performed with the INNO-LIPA test. A nested PCR followed by sequencing was used to compare different results.
Similar results were found for the two types of extraction, with an increase of sensitivity after automated extraction. Among the nine patients with a negative result with the HC2 test, seven had a negative result in INNO-LIPA and two a positive result (one untypable and one HPV66). On the 23 patients with a positive result with the HC2 test, 17 are consistent results. The six discordant results include one negative, one HPV54, two untypable HPV and two HPV53. The overall concordance between the HC2 and INNO-LIPA tests is 81% with a kappa test of 0.79.
Coupled with an automated extraction, the test INNO-LIPA confirmed its high sensitivity for detecting and typing HPV in the EasyFix media Labonord, especially in the presence of multiple genotypes. This typing systematic approach is becoming increasingly relevant in the context of HPV vaccination.
本研究的目的是通过比较QIAamp DNA Blood Mini Kit(Qiagen)提取试剂盒和自动化方法Nuclisens easyMAG(生物梅里埃公司),验证在Liquid cytology media EasyFix Labonord上使用INNO-LIPA HPV基因分型检测试剂盒(Innogenetics)的效果。
通过Hybrid Capture 2(HC2)Digene(Qiagen)技术对32个样本进行分型。通过手动或自动提取方法提取DNA,并通过PCR“HLA”进行质量控制。使用INNO-LIPA检测试剂盒进行分型。采用巢式PCR后测序来比较不同结果。
两种提取方法得到了相似的结果,自动提取后灵敏度有所提高。在HC2检测结果为阴性的9例患者中,7例在INNO-LIPA检测中结果为阴性,2例为阳性(1例无法分型,1例为HPV66)。在HC2检测结果为阳性的23例患者中,17例结果一致。6例不一致的结果包括1例阴性、1例HPV54、2例无法分型的HPV和2例HPV53。HC2检测与INNO-LIPA检测的总体一致性为81%,kappa检验值为0.79。
结合自动化提取,INNO-LIPA检测试剂盒在Labonord EasyFix培养基中检测和分型HPV时证实了其高灵敏度,尤其是在存在多种基因型的情况下。这种分型系统方法在HPV疫苗接种背景下变得越来越重要。