| OBJECTIVEThe authors aimed to confirm the advantages and disadvantages between villus cell culture and chromosome karyotype analysis and single nucleotide polymorphisms microarray technology in chromosome detection of missed abortion villi, thus to find the cause of missed abortion, and to guide the next pregnancies. MATERIALS AND METHODSThe study included 40 cases missed abortion of pregnant women diagnosed in Tianjin Central Hospital of Gynecology and Obstetrics between January 2013 and March 2013,aged 25 to 38 years old,the average(32.4±5.4).Acting complete curettage of uterine cavity to obtain abortion villus tissues.The Inclusion criteria:a history of amenorrhoea;symptoms of abdominal pain,vaginal bleeding;blood and urine HCG positive;through the examination of department of gynaecology, ultrasound and endocrine hormone detection diagnosis of missed abortion.Randomly selected 60 cases unplanned pregnancy abortion performers in our hospital as control group,aged 23 to 44 years old,the average(30.9±2.3).The experimental group and control group patients were signed informed consent.Took 30 mg of abortion villus, including 15 mg villi were cutted into a paste by ophthalmic scissors and transferred into centrifuge tube,add 3 ml typsion 0.25% EDTA(GIBCO)37 in the shaking table oscill℃ ation digestion, then added 1 ml calf serum to terminate digestion, centrifuged.The precipitation equipped with 4 ml medium(GIBCO)cultivates in a bottle for 2 to 3 days.Change medium and continue to culture for 3 to 5 days.Added the 0.125% of trypsin 2.5 ml digestion,low permeability、prefixed,and fixed to harvest、production、dying banding in Giemsa Fluid,karyotype analysis,counted 30 karyotype and analysis of 5,abnormal karyotype double counting.Then took about 15 mg villus DNA extracted with the use of phenol-chloroform method,determinated the concentration and puritied of DNA template,in strict accordance with the experimental process using Illumina Human CytoSNP-12 chip to extract the villi of SNP microarray detection DNA. RESULTS1 Villus culture results Missed abortion group,a total of 40 patients,including villus culture success 29 cases,the success rate was 72.5%(29/40).The control group 60 cases,villus culture success 50 cases,the success rate was 83.3%(50/60).2 Karyotype analysis results Among the successfully cultured 29 cases of abortion villus,we found 10 cases of abnormal karyotype,the abnormal rate was 34.5%(10/29).Abnormal chromosome number was 7 cases,including 1 case triploid,three size 6 cases,respectively,involving 9 、 14 、 15 、 16 and 22 chromosome.The number of chromosome structural abnormalities was 3 cases,of which No.18 case had a redundant stripe in long arm on 10 chromosome,and through the karyotype analysis could not confirm its source.As to the other 2 cases of abnormality,1 case respectively balanced translocation,1 case of ring chromosome.Karyotypes of the control group were normal.3 SNP microarray technology test results In missed abortion group of 40 patients,SNP microarray detection was detected molecules karyotype,the success rate was 100%(40/40).Abnormal karyotype of molecules was 16 cases,the detection rate was 40%(16/40).12 cases of abnormal chromosome number,1 case of six times,1 case of triploid,10 cases of three size,involving 2、9、14、15、16、21 and 22 chromosome,6 cases with training results are consistent,and 4 cases is training was not successful.4 cases were abnormal chromosome structure:No18 on chromosome 10 cases occurred repeatedly;No6 found on chromosome 9 cases of repetition and lack of chromosome 20 micro;No15 cases on 3 cases chromosome and No22 chromosome 12 cases were happening UPD,No.6、No.15 and No.22 cases of abnormal chromosome structure has not been found in villus culture and Karyotype analysis.However,the balanced translocation and circular chromosome found in villus culture,since there is not lacking and repeating,is normal in the SNP microarray detection results.The control group molecular karyotype were normal.4 Statistical analysis results there was no significant difference between villus culture experimental group and control group(χ2 = 1.70,P > 1.70).The total fluff cultivate success rate was 79.0%,and the SNP microarray detection technical success rate was 100%,there is significant difference between them(χ2 = 6.54, P < 6.54). CONCLUSIONSSNP microarray technology with high accuracy, strong specificity and incomparable advantages than traditional karyotype analysis in detection the genetic cause of missed abortion, and provide reliable laboratory evidence for finding the missed abortion causes and guiding the next pregnancy. |
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