Tognon Mauro, Tagliapietra Andrea, Magagnoli Federica, Mazziotta Chiara, Oton-Gonzalez Lucia, Lanzillotti Carmen, Vesce Fortunato, Contini Carlo, Rotondo John Charles, Martini Fernanda
Department of Medical Sciences, University of Ferrara, Fossato di Mortara street, 64, 44121 Ferrara, Italy.
Vaccines (Basel). 2020 Aug 25;8(3):473. doi: 10.3390/vaccines8030473.
Viral infections are considered to be risk factors for spontaneous abortion (SA). Conflicting results have been reported on the association between Human Papillomavirus (HPV) and SA. HPV DNA was investigated in matched chorionic villi tissues and peripheral blood mononuclear cells (PBMCs) from women who experienced SA ( = 80, cases) and women who underwent a voluntary interruption of pregnancy (VI; = 80, controls) by qualitative PCR and quantitative droplet digital PCR (ddPCR). Viral genotyping was performed using real-time PCR in HPV-positive samples. Specific IgG antibodies against HPV16 were investigated in sera from SA ( = 80) and VI ( = 80) females using indirect ELISA assays. None of the DNA samples from SA subjects was HPV-positive (0/80), whilst HPV DNA was detected in 2.5% of VI women ( > 0.05), with a mean viral DNA load of 7.12 copy/cell. VI samples ( = 2) were found to be positive for the HPV45 genotype. The ddPCR assay revealed a higher number of HPV-positive samples. HPV DNA was detected in 3.7% and 5% of SA and VI chorionic tissues, respectively, with mean viral DNA loads of 0.13 copy/cell in SA and 1.79 copy/cell in VI ( >0.05) samples. All DNA samples from the PBMCs of SA and VI females tested HPV-negative by both PCR and ddPCR. The overall prevalence of serum anti-HPV16 IgG antibodies was 37.5% in SA and 30% in VI ( > 0.05) women. For the first time, HPV DNA was detected and quantitatively analyzed using ddPCR in chorionic villi tissues and PBMCs from SA and VI women. Circulating IgG antibodies against HPV16 were detected in sera from SA and VI females. Our results suggest that HPV infection in chorionic villi may be a rare event. Accordingly, it is likely that HPV has no significant role in SA.
病毒感染被认为是自然流产(SA)的危险因素。关于人乳头瘤病毒(HPV)与SA之间的关联,已有相互矛盾的报道。通过定性PCR和定量微滴式数字PCR(ddPCR),对经历自然流产的女性(n = 80,病例组)和接受人工流产的女性(n = 80,对照组)的配对绒毛组织和外周血单个核细胞(PBMC)中的HPV DNA进行了研究。在HPV阳性样本中,使用实时PCR进行病毒基因分型。采用间接ELISA法,对SA组(n = 80)和人工流产组(n = 80)女性血清中针对HPV16的特异性IgG抗体进行了研究。SA组受试者的DNA样本均未检测出HPV阳性(0/80),而人工流产组女性中有2.5%检测出HPV DNA(P>0.05),平均病毒DNA载量为7.12拷贝/细胞。人工流产组样本(n = 2)被发现HPV45基因型呈阳性。ddPCR检测显示HPV阳性样本数量更多。SA组和人工流产组绒毛组织中分别有3.7%和5%检测出HPV DNA,SA组平均病毒DNA载量为0.13拷贝/细胞,人工流产组为1.79拷贝/细胞(P>0.05)。SA组和人工流产组女性PBMC的所有DNA样本经PCR和ddPCR检测均为HPV阴性。SA组女性血清抗HPV16 IgG抗体的总体患病率为37.5%,人工流产组为30%(P>0.05)。首次使用ddPCR对SA组和人工流产组女性的绒毛组织和PBMC中的HPV DNA进行了检测和定量分析。在SA组和人工流产组女性血清中检测到了针对HPV16的循环IgG抗体。我们的结果表明,绒毛组织中的HPV感染可能是罕见事件。因此,HPV在SA中可能没有显著作用。