临床实践中对前病毒DNA进行基因分型嗜性检测的表现：DIVA研究组的结果
Performance of genotypic tropism testing on proviral DNA in clinical practice: results from the DIVA study group.
作者信息
Svicher Valentina, Alteri Claudia, Montano Marco, D'Arrigo Roberta, Andreoni Massimo, Angarano Gioacchino, Antinori Andrea, Antonelli Guido, Allice Tiziano, Bagnarelli Patrizia, Baldanti Fausto, Bertoli Ada, Borderi Marco, Boeri Enzo, Bon Isabella, Bruzzone Bianca, Callegaro Anna Paola, Capobianchi Maria Rosaria, Carosi Giampiero, Cauda Roberto, Ceccherini-Silberstein Francesca, Clementi Massimo, Chirianni Antonio, Colafigli Manuela, D'Arminio Monforte Antonella, De Luca Andrea, Di Biagio Antonio, Di Nicuolo Giuseppe, Di Perri Giovanni, Di Pietro Massimo, Di Santo Fabiola, Fabeni Lavinia, Fadda Giovanni, Galli Massimo, Gennari William, Ghisetti Valeria, Giacometti Andrea, Gori Caterina, Gori Andrea, Gulminetti Roberto, Leoncini Francesco, Maffongelli Gaetano, Maggiolo Franco, Manca Giuseppe, Gargiulo Franco, Martinelli Canio, Maserati Renato, Mazzotta Francesco, Meini Genny, Micheli Valeria, Monno Laura, Mussini Cristina, Narciso Pasquale, Nozza Silvia, Paolucci Stefania, Pal Giorgio, Parisi Saverio, Parruti Giustino, Pignataro Angela Rosa, Pollicita Michela, Quirino Tiziana, Re Maria Carla, Rizzardini Giuliano, Santangelo Rosaria, Scaggiante Renzo, Sterrantino Gaetana, Turriziani Ombretta, Vatteroni Maria Linda, Vecchi Laura, Viscoli Claudio, Vullo Vincenzo, Zazzi Maurizio, Lazzarini Adriano, Perno Carlo Federico
机构信息
Department of Experimental Medicine, University of Rome, Roma, Italy.
出版信息
New Microbiol. 2012 Jan;35(1):17-25. Epub 2012 Jan 10.
OBJECTIVE
The DIVA study is aimed at setting up a standardized genotypic tropism-testing on proviral-DNA for the routine clinical diagnostic-laboratory.
METHODS
Twelve local centres and 5 reference centres (previously cross-validated) were identified. For inter-center validation-procedure, 60 peripheral-blood mononuclear cells (PBMCs) aliquots from 45 HAART-treated patients were randomly chosen for population V3 sequencing on proviral-DNA at local HIV centre and at reference-laboratory. Viral tropism was predicted by Geno2Pheno algorithm (False Positive Rate [FPR] = 20%) as proposed by the European-Guidelines. Quantification of total HIV-1 DNA was based on a method described by Viard (2004).
RESULTS
Quantification of HIV-1 DNA was available for 35/45 (77.8%) samples, and gave a median value of 598 (IQR:252- 1,203) copies/10 PBMCs. A total of 56/60 (93.3%) samples were successfully amplified by both the reference and the local virological centers. The overall concordance of tropism prediction between local and reference centers was 54/56 (96.4%). Results of tropism prediction by local centers were: 33/54 (61.1%) R5 and 21/54 (38.9%) X4/DM.
CONCLUSION
There was high concordance in the genotypic tropism prediction based on proviral DNA among different virological centers throughout Italy. Our results are in line with other European studies, and support the use of genotypic tropism testing on proviral DNA in patients with suppressed plasma HIV-1 RNA candidate to CCR5-antagonist treatment.
目的
DIVA研究旨在建立一种针对前病毒DNA的标准化基因分型嗜性检测方法，用于常规临床诊断实验室。
方法
确定了12个本地中心和5个参考中心（先前已进行交叉验证）。对于中心间验证程序，从45例接受高效抗逆转录病毒治疗（HAART）的患者中随机选取60份外周血单个核细胞（PBMC）样本，分别在本地HIV中心和参考实验室进行前病毒DNA的V3区群体测序。按照欧洲指南的建议，采用Geno2Pheno算法（假阳性率[FPR]=20%）预测病毒嗜性。HIV-1总DNA的定量基于Viard（2004年）描述的方法。
结果
45份样本中有35份（77.8%）获得了HIV-1 DNA定量结果，中位数为598（四分位间距：252-1203）拷贝/10个PBMC。参考中心和本地病毒学中心均成功扩增了56/60（93.3%）的样本。本地中心和参考中心之间嗜性预测的总体一致性为54/56（96.4%）。本地中心嗜性预测结果为：33/54（61.1%）为R5型，21/54（38.9%）为X4/DM型。
结论
意大利不同病毒学中心基于前病毒DNA的基因分型嗜性预测具有高度一致性。我们的结果与其他欧洲研究一致，支持对血浆HIV-1 RNA被抑制且可能接受CCR5拮抗剂治疗的患者进行前病毒DNA基因分型嗜性检测。
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