Gene detection kit of endometriosis Pap smear sample and method for judging deterioration degree of endometriosis
A technique for Pap smear and endometriosis, applied in the fields of genomics, biochemical equipment and methods, microbial determination/inspection, etc.
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Embodiment 1
[0038] 1. Establishment of an ethnic group experiment model
[0039] This group experiment model includes an experimental group and a control group. The experimental group included 218 subjects, all of whom were diagnosed with endometriosis after autopsy or endoscopic pathological diagnosis at the China Medical University Hospital (CMUH) in Taiwan, China. The patient's reproductive status associated with the disease was confirmed by clinical reports. According to the American Society for Reproductive Medicine (ASRM), the severity of endometriosis is divided into: stage 1, mild; stage 2, mild; stage 3, moderate; stage 4, severe. The control group consisted of 202 healthy females of the same age as the experimental group, all of whom underwent routine physical examination in the same hospital. Even if the health examination was normal, those with ovarian cysts or endometriosis-related symptoms detected by ultrasound were excluded from the control group. The above-mentioned ex...
Embodiment 2
[0054] Example 2. Establishment of a predictive model for assessing the severity of endometriosis
[0055] 1. Risk analysis of cancer-related SNPs (MiRSNPs) in miRNA genes
[0056] In the HapMap database (web site: www.hapmap.org), the minor allele frequency ( The six non-redundant SNPs with minor allele frequencies (MAF) exceeding 4%, also known as MiRSNPs. These MiRSNPs as risk factors for different cancers are listed in Table 1. These MiRSNPs are located in immature or mature miRNAs, interfering with their stability and folded configuration. The results of this example show that the genetic variation of miR-100 gene at rs1834306 (p=3.5×10 -3 , OR: 1.64; 95% CI: 1.24-2.17), and the gene variation of miR-196a2 gene at rs116l4913 (p=3.5x10 -3 , OR: 1.65; 95% CI: 1.24-2.19), both of which are associated with the risk of endometriosis, as shown in Table 2. The C allele at rs116l4913 has a dominant effect on the susceptibility of endometriosis; while patients with CC genotyp...
Embodiment 3
[0102] 1. Sample source
[0103] The conditions of 23 endometriosis patients diagnosed by China Medical University Hospital (CMUH) were also compared with the clinical reports of laparotomy or laparoscopic surgery. According to the definition of American Society of Reproductive Medicine (ASRM), the severity of endometriosis is divided into four stages. All healthy samples obtained from healthy women have undergone regular physiological tests at Kaohsiung Veterans General Hospital. In this example, informed consent was obtained from each participant. All samples were placed on smears, dehydrated with ethanol, and stored in freeze-dried form for the following experiments.
[0104] 2. Immunofluorescence Staining
[0105] The above smears were soaked in 100% ethanol for 5 minutes, 95% ethanol for 3 minutes, 75% ethanol for 3 minutes, 50% ethanol for 3 minutes and water for 5 minutes. Next, the above smear was incubated with blocking buffer (containing 0.1% Triton X-100 and 1% ...
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