ABSTRACT
<p><b>OBJECTIVE</b>To evaluate multiplex ligation-dependent probe amplification (MLPA) assay detection in analysis of chromosome 22q11.2 microdeletion.</p><p><b>METHODS</b>Between March 2008 and September 2009, thirty-two patients including 10 males and 16 females aged between years (3.6±3.1) were selected and evaluated by history, physical examination and medical records. Of these patients, sixteen patients who were previous diagnostic as 22q11.2 microdeletion were in positive control group, the other 16 healthy children were in negative control group. All the patients were detected by MLPA and fluorescence in situ hybridization (FISH) for the presence of a 22q11.2 microdeletion after informed consent. Diagnostic efficacy was assessed by sensitivity, specificity and Kappa analysis.</p><p><b>RESULTS</b>We have applied the two assays of detection of chromosome 22q11.2 microdeletion in 32 patients. Sixteen patients in positive control group were found to have a 22q11.2 deletion and, with the deletion size of 3-Mb. However, as expected, chromosome 22q11.2 deletion was not found in negative control group. The MLPA results were in good agreement with that by FISH. Therefore, MLPA has high sensitivity and specificity.</p><p><b>CONCLUSION</b>MLPA is a rapid, reliable, high-throughput and relatively economical alternative to FISH technology for the diagnosis of 22q11.2 microdeletion. It can provide reliable and helpful information for clinical diagnosis of 22q11.2 microdeletion syndrome.</p>
Subject(s)
Child, Preschool , Female , Humans , Male , Chromosome Deletion , Chromosomes, Human, Pair 22 , In Situ Hybridization, Fluorescence , Methods , Nucleic Acid Amplification Techniques , Methods , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To analyze the aberrant der(X) chromosome using conventional and molecular cytogenetic approaches in a fetus of second trimester and to discuss its clinical effect.</p><p><b>METHODS</b>Conventional cytogenetic procedures (GTG and CBG banding) were performed on cultured amniotic fluid cells. Three-color fluorescence in situ hybridization (FISH) consisting of X chromosome enumeration probes(CEPX), CEPY and Tel Xp/Yp was further performed to study the aberrant der(X) chromosome.</p><p><b>RESULTS</b>Der(X) was a rare X/Y translocation. The final karyotypes of the fetus was designated as: 46,X,der(X)t(X;Y)(p22.3;q11.2). ish der(X)t(X;Y)(p22.3;q11.2)(X/Ypter-, DXZ1+, DYZ1+)mat.</p><p><b>CONCLUSION</b>The combination of FISH and conventional cytogenetic techniques is a powerful tool to determine derivative chromosome and to offer an accurate genetic counseling. Identification of Xp; Yq rearrangement can help estimate the risk of fetus abnormalities and give a more precise prognosis.</p>
Subject(s)
Adult , Female , Humans , Pregnancy , Amniocentesis , Methods , Amniotic Fluid , Cell Biology , Chromosome Aberrations , Chromosome Banding , Methods , Chromosomes, Human, X , Cytogenetic Analysis , Methods , Fetus , Congenital Abnormalities , Genetic Counseling , Methods , In Situ Hybridization, Fluorescence , Methods , Pregnancy Trimester, SecondABSTRACT
<p><b>OBJECTIVE</b>To analyze the numerical aberration rate of X, Y and chromosome 18 in sperms from an oligozoospermic male with mosaic trisomy 18 and to perform preimplantation genetic diagnosis (PGD) for the couple.</p><p><b>METHODS</b>G-banding and fluorescence in situ hybridization (FISH) were performed on metaphase chromosome. Sperm was analyzed in three-color FISH with a probe mixture containing CEP18, CEPY and Tel Xq/Yq. A healthy man with normal semen parameters was used as control.</p><p><b>RESULTS</b>Significant difference in the rates of disomy for chromosome 18 (0.63% vs. 0.16%) and the gonosomes (0.945% vs. 0.35%) and diploidy (0.87% vs. 0.31%) was found in the spermatozoa between the patient and the control. After four embryos were biopsied in one PGD cycle, two embryos with XY1818 and XX1818 were selected for implanting and clinical pregnancy was ongoing.</p><p><b>CONCLUSION</b>Sperm-FISH allows further understanding of aneuploidy rate and accurate genetic counseling. FISHPGD was effective for patient with mosaic trisomy 18.</p>
Subject(s)
Humans , Male , Aneuploidy , Chromosomes, Human, Pair 18 , Chromosomes, Human, Y , Genetics , Diploidy , In Situ Hybridization, Fluorescence , Infertility, Male , Genetics , Oligospermia , Diagnosis , Genetics , Preimplantation Diagnosis , Semen Analysis , Spermatozoa , Trisomy , Diagnosis , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the application of differential display-2PCR(DD-PCR) in research on gene expression of Candida.</p><p><b>METHODS</b>Resistance to fluconazole was induced in a Candida albicans isolate 435 from vagina by culturing in YEPD broth with increasing fluconazole concentration in vitro, and the resistant isolate 435-2 (MIC=128 microg/ml ) was obtained after 80 days of incubation. Comparisons between 435 and 435-2 either in fluconazole-containing medium or in drug-free medium were performed with the modified DD-PCR including amplification with long primers, silver staining, reverse dot blot and non-radiographic labeling techniques.</p><p><b>RESULTS</b>Three differential displayed bands were found which showed high homology to alcohol dehydrogenase 1 (ADH1), TOP2 and CDR1, respectively. The up-regulating expression of ADH1 and CDR1 associated with fluconazole resistance was further identified by RT-PCR.</p><p><b>CONCLUSION</b>The up-regulating expression of ADH1 and CDR1 was associated with fluconazole resistance in Candida albicans, ADH1 might be a candidate of novel fluconazole resistant gene.</p>