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目的探讨高级别宫颈鳞状上皮内瘤变(HSIL)锥切手术前后阴道菌群构成的差异。方法选择2015年12月至2016年8月南方医科大学珠江医院、广东省人民医院收治的HSIL患者20例,分别采集其锥切手术前与手术后3个月的阴道分泌物,同时检测高危型HPV-DNA含量(HC2法);随后对每个样本进行细菌总基因组DNA提取、16S rRNA V4区基因扩增及采用Illumina高通量测序技术对扩增的PCR产物进行测序等步骤,然后通过BIPES生物信息分析方法比较HSIL患者手术前后阴道菌群物种丰度和结构的差异。结果 HSIL患者术前高危型HPV-DNA均为阳性,术后3个月第1次复查均转阴性。HSIL患者手术前的阴道菌群以惰性乳酸杆菌、加德纳菌属、普氏菌属、奇异菌属等为优势菌属,而卷曲乳酸杆菌含量低。在锥切手术后,惰性乳酸杆菌、加德纳菌属、普氏菌属、奇异菌属等则明显降低,卷曲乳酸杆菌则有升高趋势。术后则以卷曲乳酸杆菌、脲原体、普氏菌属、肠球菌属等为优势菌属。结论高级别宫颈鳞状上皮内瘤变患者经手术去除病变后,高危型HPV含量显著减少甚至消失,其阴道菌群出现了向常见阴道优势菌属转变的趋势。提示HPV可能与阴道菌群构成密切相关且相互影响。
Objective To investigate the differences of vaginal flora between high-grade cervical squamous intraepithelial neoplasia (HSIL) conization before and after surgery. Methods From December 2015 to August 2016, 20 cases of HSIL patients admitted to Zhujiang Hospital of Southern Medical University and Guangdong Provincial People’s Hospital were collected for vaginal discharge before and 3 months after conization, HPV-DNA content (HC2 method); Then for each sample total bacterial genomic DNA extraction, 16S rRNA V4 region gene amplification and sequencing using Illumina high-throughput sequencing of the amplified PCR products and other steps, and then by BIPES Bioinformatics analysis of HSIL patients before and after vaginal flora abundance and structure differences. Results The preoperative high-risk HPV-DNA in HSIL patients were all positive. The first review at 3 months after operation was negative. Vaginal flora of patients with HSIL before surgery with lactic acid bacteria Lactobacillus, Gardnerella, Prevotella, such as genus Culex is the dominant genus, and curly Lactobacillus content is low. After conization surgery, lactic acid bacteria Lactobacillus, Gardnerella, Prevotella, and other bacteria were significantly reduced, curly Lactobacilli there is an upward trend. After surgery with curly Lactobacillus, Ureaplasma, Prevotella, Enterococcus and other dominant bacteria. Conclusion The high-risk HPV types in patients with high-grade cervical squamous intraepithelial neoplasia were significantly reduced or even disappeared after surgical removal of the lesions, and their vaginal flora tended to change to common vaginal dominant bacteria. Tip HPV may be closely related with the vaginal flora and influence each other.