Carcinoma ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Pancreatic Neoplasms ; genetics ; Spectral Karyotyping ; methods

OBJECTIVETo rapidly detect SOX2 gene using primed in situ labeling (PRINS).

METHODSHuman peripheral blood samples were cultured using an optimized method. Sequence of the SOX2 gene was amplified in situ with biotin-labeled specific primers and processed with a tyramide signal amplification (TSA) biotin system. Subsequently, fluorescence-stained signal was detected by streptavidin-Texas red. For the control group, MCF-10F cells were transfected with Lentivirus hSox2.

RESULTSBy VideoTesT-FISH software analysis, the long arm of chromosome 3 in the experimental group showed a specific red fluorescence signal, whilst the control samples showed no specific signals for SOX2. Transfected MCF-10F cells showed various efficiency of SOX2 gene integration.

CONCLUSIONPRINS utilizes a highly sensitive in situ PCR technique combined with fluorescence labeled oligodeoxynucleotides can synthesize probes in situ, thus greatly reducing the cost of probe and time for detection. It can facilitate identification and classification of induced pluripotent stem cells, and has many potential applications in this prospect.

Humans ; In Situ Hybridization, Fluorescence ; methods ; Male ; Primed In Situ Labeling ; methods ; SOXB1 Transcription Factors ; chemistry

OBJECTIVETo establish a primed in situ labeling (PRINS) technique which can be more effective in detection of single copy gene.

METHODSOn the basis of traditional PRINS, new reagents and procedures, such as TaqStart antibody, four primers of the sex determining region Y (SRY) gene and TSA(TM) Biotin System were included in detection of the SRY gene. Meanwhile, fluorescence in situ hybridization(FISH) to detect the SRY gene was used as control.

RESULTSFifty metaphases were scored. PRINS labeling showed signals for the SRY on the Y chromosome at band Yp11.3 in all metaphases. These signals were as distinct as that from results of FISH.

CONCLUSIONThis improved method is ideal for rapidly localizing single copy genes and small DNA segments. And PRINS is a cost- and time-effective alternative to FISH.

Gene Dosage ; Genes, sry ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Metaphase ; genetics ; Primed In Situ Labeling ; methods

No abstract available.
Fluorescence* ; In Situ Hybridization*

Consensus ; In Situ Hybridization, Fluorescence

OBJECTIVETo determine the value of spectral karyotyping (SKY) to identify the complex chromosome aberration.

METHODSFour cases were selected that can not be identified by standard cytogenetic techniques. The chromosome specimens were detected by the routine SKY method, and the results were analyzed by the SKY View software.

RESULTSBy using SKY a case of complex chromosome rearrangements and two cases of chromosome duplication were identified. However it could not identify the chromosome inversion and the breakpoint of chromosome aberration.

CONCLUSIONSKY may be a valuable tool in identification of complex chromosome translocation, rearrangement, minute aberration and unknown derivative chromosomes. Though SKY can not replace the standard cytogenetic techniques, but it will be the benefit supplementary.

Adult ; Chromosome Aberrations ; Chromosome Banding ; Chromosome Disorders ; diagnosis ; genetics ; Female ; Gene Duplication ; Humans ; In Situ Hybridization, Fluorescence ; Infant ; Male ; Reproducibility of Results ; Sensitivity and Specificity ; Spectral Karyotyping ; methods ; Translocation, Genetic

OBJECTIVEComprehensive use of molecular cytogenetic techniques for the detection of 1 case of small chromosome translocation.

METHODSFollowing conventional chromosome preparation, G-banding karyotype analysis, spectral karyotyping (SKY), whole chromosome painting, two-color fluorescence in situ hybridization (FISH) and subtelomeric probe FISH were performed.

RESULTSG-banded karyotype was 46, XX, ?(22q11.3), SKY karyotype analysis was 46, XX, der (4)t(4;6) and found no abnormalities on chromosome 22, staining signal was not found with any abnormalities on chromosome 6. Two-color FISH indicated a chromosomal translocation segment of 22q13.3 to one end of the short arm of chromosome 4. Subtelomeric FISH probe showed the end of the long arm of chromosome 22 and the end of the short arm of chromosome 4 reciprocal translocation. High resolution G-banding and FISH result indicated 46, XX, t(4;22)(p15.3;q13.2).

CONCLUSIONThe testing of small chromosomal translocation should be combined with clinical information and integrated use of molecular cytogenetic techniques to improve the accuracy of diagnosis of chromosomal diseases.

Adult ; Chromosome Banding ; Chromosomes, Human, Pair 22 ; genetics ; Chromosomes, Human, Pair 4 ; genetics ; Cytogenetic Analysis ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Spectral Karyotyping ; Translocation, Genetic ; genetics

OBJECTIVE: Increasingly it is being recognized that genetic factors play a significant role in causing malformation. There are many available prenatal diagnostic methods including cytogenetic karyotyping using amniocentesis and cordocentesis, fluorescence in situ hybridization(FISH), and primed in situ labelling(PRINS). Our purpose was to attempt to discuss the clinical use of cytogenetic karyotyping, FISH, and PRINS. METHODS: We conducted 222 cases of cytogenetic karyotyping using amniocentesis and cordocentesis, l0 cases of FISH, and 10 cases of PRINS from January 1996 to July 1998 at Ewha Womans University Mokdong Hospital. Age distribution, chromosomal abnormalities by age group, indication, karyotype, and baby outcomes were performed. RESULTS: Overall incidence of chromosomal abnormalities was 7.7%(17cases) and chromosomal abnormalities were most frequently noted in 30-34 year old women and 35-39 year old women(2.3%, respectively). Among 222 cases, 25-29 year old women were highest(30.2%). Chromosomal abnormalities among cytogenetic karyotyping cases were Down syndrome, Edward syndrome, Patau syndrome, Deletion(8), Inversion(9), etc. The 5 cases of healthy baby among chromosomal abnormalities were delevered. Among 213 cases of karyotyping using amniocentesis, abnormal karyotyping cases were 15 cases. Among 15 cases, 8 cases were terminated and 5 cases of healthy baby were delivered. Among 9 cases of karyotyping using cordocentesis, 2 cases of chromosomal abnormalities(Edward, Down syndrome) were found and 3 cases healthy baby were delivered. Among 10 cases of FISH results, 6 case of FISH results were the same with G-banding and were different from G-banding. Among 10 cases of PRINS results, we got the PRINS results from 7 cases. CONCLUSION: It is concluded that cytogenetic karyotyping, FISH, and PRINS are very useful to detect chromosomal abnormalities.
Age Distribution ; Amniocentesis ; Chromosome Aberrations ; Cordocentesis ; Cytogenetics* ; Down Syndrome ; Female ; Fluorescence* ; Humans ; In Situ Hybridization* ; Incidence ; Karyotype ; Karyotyping* ; Prenatal Diagnosis* ; Primed In Situ Labeling

No abstract available.
Humans ; In Situ Hybridization, Fluorescence* ; Infant, Newborn*

Humans ; In Situ Hybridization, Fluorescence ; methods ; Paraffin