Preliminary research for the determination of HPV E6/E7 mRNA by nucleic acid molecular hybridization-flow cytometry

doi:10.3969/j.issn.1673-8640.2023.01.009

Abstract

Abstract:

Objective To investigate the reliability and application value of determining human papillomavirus（HPV） E6/E7 mRNA by nucleic acid molecular hybridization-flow cytometry. Methods Totally，361 samples of cervical epithelial exfoliated cells collected from gynecological departments of Guangdong Maternal and Child Health Hospital and the First Affiliated Hospital of Sun Yat-sen University from October 2018 to October 2019 were collected，and thinprep cytologic test（TCT），HPV DNA，biopsy histopathology and HPV E6/E7 mRNA determinations were performed. Results The proportion of positive HPV E6/E7 mRNA results was increased with the severity of pathological biopsy results. Compared with HPV DNA，HPV E6/E7 mRNA had higher specificity（72.73%）and positive predictive value（47.06%）. According to HPV E6/E7 mRNA results，the accuracy of colposcopy shunt of atypical squamous cell of undetermined significance（ASCUS） in abnormal TCT results was 66%. Conclusions HPV E6/E7 mRNA has high consistency with histopathological examination，and the accuracy of colposcopy shunt，specificity and positive predictive value for high-grade lesions are high.

Key words: Human papillomavirus, E6/E7 mRNA, Cervical cancer, Nucleic acid molecular hybridization, Flow cytometry

CLC Number:

R373.9

ZHANG Yaxu, CUI Yanwei, LIU Ziyan. Preliminary research for the determination of HPV E6/E7 mRNA by nucleic acid molecular hybridization-flow cytometry[J]. Laboratory Medicine, 2023, 38(1): 46-50.

Figures/Tables 5

References 15

[1]	PLUMMER M, DE MARTEL C, VIGNAT J, et al. Global burden of cancers attributable to infections in 2012:a synthetic analysis[J]. Lancet Glob Health, 2016, 4(9):e609-e616. DOI URL
[2]	吕秀芳, 孔令非. 人乳头状瘤病毒E6/E7 mRNA研究进展[J]. 中华实用诊断与治疗杂志, 2016, 30(2):111-113.
[3]	OUH Y T, LEE J K. Proposal for cervical cancer screening in the era of HPV vaccination[J]. Obstet Gynecol Sci, 2018, 61(3):298-308. DOI URL
[4]	苏瑛, 谢婷婷, 于月成. HPV E6/E7 mRNA在宫颈癌中的应用进展[J]. 中国生育健康杂志, 2018, 29(1):92-94.
[5]	叶丽君, 喻长法, 蔡莎莎, 等. 高危型人乳头瘤病毒E6/E7 mRNA联合液基细胞学检查在宫颈病变诊断中的应用[J]. 中国慢性病预防与控制, 2017, 25(7):499-501.
[6]	LIU S, MINAGUCHI T, LACHKAR B, et al. Separate analysis of human papillomavirus E6 and E7 messenger RNAs to predict cervical neoplasia progression[J]. PLoS One, 2018, 13(2):e0193061. DOI URL
[7]	黄文林. 分子病毒学[M]. 北京: 人民卫生出版社, 2016.
[8]	李晓林, 袁文娟, 陈龙, 等. 高危型HPV E6/E7 mRNA检测在宫颈机会性筛查中应用价值研究[J]. 中华肿瘤防治杂志, 2017, 24(4):217-222.
[9]	LU Z, CHEN H, ZHENG X M, et al. Expression and clinical significance of high risk human papillomavirus and invasive gene in cervical carcinoma[J]. Asian Pac J Trop Med, 2017, 10(2):195-200. DOI PMID
[10]	PRUSKI D, MILLERT-KALINSKA S, LEWEK A, et al. Sensitivity and specificity of HR HPV E6/E7 mRNA test in detecting cervical squamous intraepithelial lesion and cervical cancer[J]. Ginekol Pol, 2019, 90(2):66-71. DOI PMID
[11]	REBOLJ M, PREISLER S, EJEGOD D M, et al. Disagreement between human papillomavirus assays:an unexpected challenge for the choice of an assay in primary cervical screening[J]. PLoS One, 2014, 9(1):e86835. DOI URL
[12]	梁海燕, 陈晓红. HPV DNA和E6/E7 mRNA检测对宫颈病变诊断价值比较研究[J]. 中国医药科学, 2017, 7(12):19-22.
[13]	QING W, CAIYING Z, LIMEI C, et al. Clinical value of human papillomavirus E6/E7 mRNA testing in patients with atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesion[J]. 生殖与发育医学(英文), 2018, 2(3):157-161.
[14]	DABESKI D, DUVLIS S, BASHESKA N, et al. Comparison between HPV DNA testing and HPV E6/E7 mRNA testing in women with squamous cell abnormalities of the uterine cervix[J]. Pril(Makedon Akad Nauk Umet Odd Med Nauki), 2019, 40(1):51-58.
[15]	PANDEY M, BHOSALE P G, MAHIMKAR M B. Detection of HPV E6/E7 mRNA in clinical samples using RNA in situ hybridization[J]. Methods Mol Biol, 2018, 1726:167-175. DOI PMID

CIN分级	例数	E6/E7 mRNA阴性/例	E6/E7 mRNA阳性/例	E6/E7 mRNA阳性率/%
炎症等	39	32	7	17.95
CIN 1级（或LSIL）^①	66	45	21	31.82
CIN 2级	21	13	8	38.10
CIN 2/3级（或HSIL）^②	23	14	9	39.13
CIN≥3级	9	1	8	88.89
合计	158	105	53

CIN分级	例数	E6/E7 mRNA阴性/例	E6/E7 mRNA阳性/例	E6/E7 mRNA阳性率/%
炎症等	39	32	7	17.95
CIN 1级（或LSIL）^①	66	45	21	31.82
CIN 2级	21	13	8	38.10
CIN 2/3级（或HSIL）^②	23	14	9	39.13
CIN≥3级	9	1	8	88.89
合计	158	105	53

项目	CIN≥2级/例	CIN≤1级/例
HPV E6/E7 mRNA
阳性	24	27
阴性	28	72
HPV DNA
阳性	51	87
阴性	1	12

项目	CIN≥2级/例	CIN≤1级/例
HPV E6/E7 mRNA
阳性	24	27
阴性	28	72
HPV DNA
阳性	51	87
阴性	1	12

TCT结果	例数	HPV E6/E7 mRNA阳性			HPV E6/E7 mRNA阴性
		样本数/例	组织病理学检查		样本数/例	组织病理学检查
		样本数/例	CIN≤1级或LSIL/例	CIN≥2级/例	样本数/例	CIN≤1级或LSIL/例	CIN≥2级/例
NILM	34	9	8	1	25	22	3
ASCUS	50	18	10	8	32	25	7
ASCH	5	1	1	0	4	1	3
LSIL	48	15	7	8	33	27	6
HSIL	19	9	1	8	10	2	8
AGC	1	0	0	0	1	0	1
合计	157	52	27	25	105	77	28