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正常宫颈细胞学女性的 HPV 分子检测和 FT-IR 光谱分析。

Molecular detection of HPV and FT-IR spectroscopy analysis in women with normal cervical cytology.

机构信息

Laboratório de Biologia Molecular do Câncer, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida Shishima Hifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.

Laboratório de Nanosensores, Universidade do Vale do Paraíba, UNIVAP, Instituto de Pesquisa e Desenvolvimento, Avenida ShishimaHifumi 2911, Urbanova, São José dos Campos, 12244-000, São Paulo, SP, Brazil.

出版信息

Photodiagnosis Photodyn Ther. 2020 Mar;29:101592. doi: 10.1016/j.pdpdt.2019.101592. Epub 2019 Nov 6.

Abstract

BACKGROUND

The increase in the incidence of Cervical Cancer in the female population worldwide has been an issue that deserves further attention from the scientific community. Several studies have already proven the relationship of its development with the molecular mechanisms that Human Papillomavirus (HPV) induces in cervical cells. The gene amplification provided by molecular biology techniques has been used as the gold standard diagnostic method of this virus because of its high specificity and sensitivity.However, the high investments associated with the acquisition of reagents, equipment and labor demonstrate the need for the development of more accessible techniques that present the same accuracy. FT-IR spectroscopy has been studied as an inexpensive and easily accessible technology that can provide the differentiation of malignant and benign cells. This study aimed to demonstrate the effectiveness and sensitivity of molecular analysis by PCR in relation to cytological analysis and to evaluate the sensitivity of FT-IR spectroscopy in the diagnosis of HPV for cervical cancer prevention.

METHODS

Cervical fluid samples obtained from 50 patients with absence of cellular lesion by cytological analysis were analyzed by molecular and spectroscopic analyzes. Oncotic colpocitology analysis was performed by the Papanicolaou staining, amplification of the L1 viral gene by PCR was performed using primers MY09 and MY11 and biochemical analysis of the fluids by FT-IR was performed using the Spectrum 400 system equipped with a microscope.

RESULTS

Of the 50 patients without evident morphological alteration of the cells, seven were diagnosed by molecular analysis as positive for presence of HPV. Principal component analysis of spectroscopy was not able to separate the negative samples from the HPV positive samples and, therefore, did not present as an effective diagnostic technique.

CONCLUSIONS

We highlight the efficacy, sensitivity and specificity of molecular biology by PCR in the identification of the virus and we emphasize that more studies should be used for the application of FT-IR spectroscopy in the diagnosis of this infection and its application in the prevention of cervical cancer.

摘要

背景

全球女性宫颈癌发病率的上升是科学界值得进一步关注的问题。已有多项研究证明,宫颈癌的发展与 HPV 诱导宫颈细胞的分子机制有关。分子生物学技术提供的基因扩增已被用作该病毒的金标准诊断方法,因为它具有高特异性和灵敏度。然而,与试剂、设备和劳动力获取相关的高投资表明,需要开发更易获得的技术,这些技术具有相同的准确性。傅里叶变换红外(FT-IR)光谱已被研究为一种廉价且易于获取的技术,它可以提供恶性和良性细胞的区分。本研究旨在证明分子分析 PCR 的有效性和敏感性与细胞学分析的关系,并评估 FT-IR 光谱在 HPV 诊断中的敏感性,以预防宫颈癌。

方法

对 50 例细胞学分析未见细胞病变的宫颈液样本进行分子和光谱分析。采用巴氏染色进行渗出性 colpocitology 分析,采用引物 MY09 和 MY11 对 L1 病毒基因进行扩增,采用配备显微镜的 Spectrum 400 系统对液体制备进行生化分析。

结果

在 50 例细胞形态无明显改变的患者中,有 7 例经分子分析诊断为 HPV 阳性。光谱主成分分析不能将阴性样本与 HPV 阳性样本分开,因此不能作为有效的诊断技术。

结论

我们强调了 PCR 分子生物学在识别病毒方面的功效、敏感性和特异性,并强调应更多地开展研究,以将 FT-IR 光谱应用于该感染的诊断及其在宫颈癌预防中的应用。

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