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OBJECTIVE@#To establish the rapid PCR amplification program and system and to verify the technical indexes.@*METHODS@#PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed.@*RESULTS@#The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate.@*CONCLUSION@#The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
Subject(s)
Humans , Electrophoresis, Capillary , Fluorescence , Genetics, Population , Genotype , Microsatellite Repeats , Multiplex Polymerase Chain Reaction/methods , Sensitivity and SpecificityABSTRACT
Background Surgical induced astigmatism induced by a 5.5mm of superior scleral tunnel incision is a key cause of influencing vision outcome in the high myopic eyes after implantation of Artisan phakic intraocular lens(IOL).Objective This study was to evaluate the surgical induced astigmatism through a 5.5mm superior scleral tunnel incision during the implantation of Artisan phakic IOL.Methods This was a case observational study.The clinical data of 202 eyes of 111 patients with Artisan IOL implantation for high myopia from October 2004 through October 2008 was collected.The patients were followed-up for 12 months.The uncorrected visual acuity(UCVA),best spectacle-corrected visual acuity(BCVA),spherical equivalent(SE),cylinder equivalent(CE) and cylinder axis were examined before surgery and postoperative 1,3,6,12 months.The patients were divided into with rule group and against rule group according to the cylinder axis.Surgery-induced astigmatism value was calculated using Holladay vector analysis formula.This clinical trial was approved by the Ethics Committee of Guangdong General Hospital.Written informed consent was obtained from each patient prior to the surgery.Results The UCVA of 195 eyes(94.1%) was 0.5 or better in postoperative 12 months,and that of 172 eyes(85.1%) showed the improvement of BCVA during the follow-up duration.The mean CE values were reduced in 1,3,6,12 months after surgery in comparison with before surgery(t=-4.30,P=0.00;t=-2.27,P=0.01;t=-2.04,P=0.04;t=-2.79,P=0.01).Vector analysis of total surgically induced astigmatism revealed that the mean incision-induced astigmatism value was +1.94D,+2.26D,+2.29D,+2.25D at 171°,170°,181°,175° in 1,3,6,12 months after surgery.The surgery-induced astigmatism was(+1.97±1.84)D,(+2.25±1.75)D,(+2.27±1.76)D,(+2.24±1.75)D in with rule group at postoperative 1,3,6,12 months respectively;while those in against rule group was(+1.75±1.88)D,(+2.35±1.74)D,(+2.38±1.76)D,(+2.34±1.74)D respectively.No significant differences were found between the two groups(t=0.54,-0.29,-0.27,-0.29;P=0.59,0.78,0.79,0.78).Conclusion Implantation of Artisan IOL is an effective approach for correction of high myopia.A 5.5mm superior scleral tunnel incision in operation induces a +2.25D astigmatism at 175° meridian.
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<p><b>BACKGROUND</b>To investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.</p><p><b>METHODS</b>The restriction-fragment-length-polymorphism (RFLP) assay for HBV DNA sequence determination at the codon 528 and 552 in the HBV polymerase gene associated with lamivudine resistance in vitro. HBV DNA samples extracted from sera of 240 patients were subjected to PCR amplification with primer pairs F2/R2 (552), F3/R2 (528). Each PCR product was digested with Nde I or Nla III.</p><p><b>RESULTS</b>Serum HBV DNA mutation was found in 51/240 patients (38/51M552V, 26/38L528M, 13/51M552I) after therapy for 52 weeks. DNA sequence analysis was performed on samples of 3 patients, and the results were consistent with those of RFLP assay.</p><p><b>CONCLUSION</b>The RFLP assay was able to detect the mutation of HBV DNA at codon 552 and 528 which are the principal site of HBV DNA resistant to lamivudine. The specific PCR method for HBV DNA mutation is rapid, simple and specific.</p>