This is a brief introduction to the technique and principle of Array Comparative Genomic Hybridization. Array CGH is a powerful tool for genetic testing and has been enormously useful in cancer cytogenetics, prenatal genetic testing etc.
Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells
A micro-array is a tool for analyzing gene expression that consists of a small membrane or glass slide containing samples of many genes arranged in a regular pattern.
This was made by me while I was in Masters. I have made few animations. I hope it makes understanding better.
The content is made by searching through internet and referencing books. I do not claim any content in whole presentation except the animations made on the subject.
complete Single Nucleotide Polymorphiitsm Detection methods with Advance techniques with its applications
Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
Heteroduplex Analysis (HA)
Denaturing High Performance Liquid Chromatography (DHPLC)
UNG-Mediated T-Sequencing
RNA-Mediated Finger printing with MALDI MS Detection
Sequencing by Hybridization
Direct DNA Sequencing
Single-feature polymorphism (SFP)
Invader probe
Allele-specific oligonucleotide probes
PCR-based methods
Allele specific primers
Sequence Polymorphism-Derived (SPD) markers
Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
Allele-specific ligation probes
What is in situ hybridization
Radioactive ISH
Fluorescent ISH
Colorimetric ISH
ISH: three variables
The sample
The probe
Optimizing ISH Detection
ISH controls
Data Analysis
Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells
A micro-array is a tool for analyzing gene expression that consists of a small membrane or glass slide containing samples of many genes arranged in a regular pattern.
This was made by me while I was in Masters. I have made few animations. I hope it makes understanding better.
The content is made by searching through internet and referencing books. I do not claim any content in whole presentation except the animations made on the subject.
complete Single Nucleotide Polymorphiitsm Detection methods with Advance techniques with its applications
Single nucleotide polymorphisms are single base variations between genomes within a species.
There are at least 10 million polymorphic sites in the human genome.
SNPs can distinguish individuals from one another
Denaturing Gradient Gel Electrophoresis
Chemical Cleavage Of Mismatch
Single-stranded Conformation Polymorphism (SSCP)
MutS Protein-binding Assays
Mismatch Repair Detection (MRD)
Heteroduplex Analysis (HA)
Denaturing High Performance Liquid Chromatography (DHPLC)
UNG-Mediated T-Sequencing
RNA-Mediated Finger printing with MALDI MS Detection
Sequencing by Hybridization
Direct DNA Sequencing
Single-feature polymorphism (SFP)
Invader probe
Allele-specific oligonucleotide probes
PCR-based methods
Allele specific primers
Sequence Polymorphism-Derived (SPD) markers
Targeting induced local lesions in genomes (TILLinG)
Minisequencing primers
Allele-specific ligation probes
What is in situ hybridization
Radioactive ISH
Fluorescent ISH
Colorimetric ISH
ISH: three variables
The sample
The probe
Optimizing ISH Detection
ISH controls
Data Analysis
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
Definition, principle, Chemical used during the process, application, advantages, and disadvantages of both techniques. along with relevant case study for better understand
Slideshow is from the University of Michigan Medical School's M1 Patients and Populations: Medical Genetics Sequence.
View additional course materials on Open.Michigan:
openmi.ch/med-M1PatientsPopulations
Gene mapping means the mapping of genes to specific locations on chromosomes.
Such maps indicates the positions of genes in the genome and also distance between them.
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Genome-wide association study (GWAS) technology has been a primary method for identifying the genes responsible for diseases and other traits for the past ten years. GWAS continues to be highly relevant as a scientific method. Over 2,000 human GWAS reports now appear in scientific journals. Our free eBook aims to explain the basic steps and concepts to complete a GWAS experiment.
A cytological technique to detect the nature of adjacent chromosomal regions by using different staining technique assisted with some pre treatment of metaphase chromosomes prepared on the slides
Deciphering DNA sequences is essential for virtually all branches of biological research. With the
advent of capillary electrophoresis (CE)-based Sanger sequencing, scientists gained the ability to
elucidate genetic information from any given biological system. This technology has become widely
adopted in laboratories around the world, yet has always been hampered by inherent limitations in
throughput, scalability, speed, and resolution that often preclude scientists from obtaining the essential
information they need for their course of study. To overcome these barriers, an entirely new technology
was required—Next-Generation Sequencing (NGS), a fundamentally different approach to sequencing
that triggered numerous ground-breaking discoveries and ignited a revolution in genomic science.
Definition, principle, Chemical used during the process, application, advantages, and disadvantages of both techniques. along with relevant case study for better understand
Slideshow is from the University of Michigan Medical School's M1 Patients and Populations: Medical Genetics Sequence.
View additional course materials on Open.Michigan:
openmi.ch/med-M1PatientsPopulations
Gene mapping means the mapping of genes to specific locations on chromosomes.
Such maps indicates the positions of genes in the genome and also distance between them.
A physical map of a chromosome or a genome that shows the physical locations of genes and other DNA sequences of interest. Physical maps are used to help scientists identify and isolate genes by positional cloning.
According to the ICSM (Intergovernmental Committee on Surveying and Mapping), there are five different types of maps: General Reference, Topographical, Thematic, Navigation Charts and Cadastral Maps and Plans.
Genome-wide association study (GWAS) technology has been a primary method for identifying the genes responsible for diseases and other traits for the past ten years. GWAS continues to be highly relevant as a scientific method. Over 2,000 human GWAS reports now appear in scientific journals. Our free eBook aims to explain the basic steps and concepts to complete a GWAS experiment.
High throughput screening is a type of assay. By this assay we can identified the target or binding site of drugs. Its mainly performed during the drug discovery process.
FDA/EMA Inspection - Regulatory Expectations in Early Phase Clinical TrialsSGS
FDA, EMA and other regulatory agencies routinely perform both announced and unannounced phase I clinical investigator inspections, in order to verify the accuracy and reliability of data that has been submitted to the agency, and to provide real-time assessment of the investigator’s conduct of the trial to ensure human safety.
Preparing for a clinical inspection involves making sure that all aspects of your clinical trial are up to code. This includes being able to provide all required documents that prove that your trial is GCP compliant.
A short summary of SGS CPU experience in ensuring site compliance prior to/during an agency inspection, as well as the importance of conducting site internal quality management is presented in this presentation.
For more information visit: https://www.sgs.com/exprimo and contact us at: clinicalresearch@sgs.com
Follow us on LinkedIn: https://www.linkedin.com/company/sgs
Medical Utopias: The Promise of Emerging TechnologiesAlex Tang
Medical utopias are often about good health, absence of suffering, and even delaying of the aging process. The last two decades have seen a tremendous increase in emerging medical technologies to achieve these utopias. The completion of the sequencing of the human genome sets the stage for the next step of genetic and molecular advances. The increase in computing power, storage capacity, connectivity, and the Internet has opened avenues of new diagnostic and therapeutic modalities. The perfecting of sustaining cell growth in vitro and cell nucleus transfer has opened the way to cloning, stem cell harvesting, and a new field of regenerative medicine. However, these emerging technologies bring with them a large number of bioethical concerns that need to be addressed. These concerns involving tissue engineering, bioelectronics, new genetics, cloning, gene therapy, germ-line genome modifications are only the tip of the iceberg. In this paper I will reflect on three areas of concern. Firstly, the emergence of the digital patient will be considered. This digital patient will be deeply formed and informed by health information technology (IT), the social media, and issues involving privacy, confidentiality and data security. Secondly, the direct to customers (DTC) genetic screening tests will be discussed. The ethical issue of buccal swabs taken at home and be tested for genetic diseases and future prediction of other illnesses which is marketed directly to the consumers will be examined. Finally, the development of new pharmaco-therapeutics will be explored. There have been changes in the way new drugs are tested and these changes do raise some ethical concerns. The examination of these ethical issues will be done in the framework of respect for autonomy, beneficence, non-maleficence, and justice.
Automated sequencing of genomes require automated gene assignment
Includes detection of open reading frames (ORFs)
Identification of the introns and exons
Gene prediction a very difficult problem in pattern recognition
Coding regions generally do not have conserved sequences
Much progress made with prokaryotic gene prediction
Eukaryotic genes more difficult to predict correctly
Applications of Bio systems Engineering (Artificial Organs)Dineesha Nipunajith
Applications Of Bio - Systems Engineering !
Artificial Heart
Artificial lungs
Artificial kidneys
Artificial nose
Artificial tongue
Advantages & Disadvantages
Health Risks
In situ hybridization methods and techniques course slides Pat Heslop-HarrisonPat (JS) Heslop-Harrison
Methods and techniques for chromosomal in situ hybridization and molecular cytogenetics. Fixations, chromosomes preparation, mostly using plant chromosomes, hybridiziation mixtures, stringency calculations and fluorescent microscopy.Trude Schwarzacher and Pat Heslop-Harrison
RAPD markers are decamer DNA fragments.
RAPD is a type of PCR reaction.
as the name suggest it is a fast method when compared to the traditional PCR medthod.
diagnosis of cancer, bioluminescent detection, diagnosis of cancer, haplotype mapping, imaging gene expression in vivo, types of cancer diagnosis method, ultrasound imaging
Ion Torrent™ semiconductor sequencing, combined with Ion AmpliSeq™ technology, provides simultaneous identification of copy number variants (CNVs), single nucleotide variants (SNVs), and small insertions and deletions (indels) from a research sample by means of a single integrated workflow. 100% of assayed CNV regions (n=34) were detected using a reference set of 31 samples with known chromosomal aberrations. Low-pass whole-genome sequencing data, with approximately 0.01x read coverage, allowed the rapid ≤10 hour analysis of aneuploidies from research samples with extremely low initial input DNA amounts—even from a single cell. Using a control set of 10 samples with known chromosomal aberrations, 100% of the copy number changes were found, ranging from gains or losses of whole chromosomes to subchromosomal alterations tens of megabases (Mb) in size. The Ion PGM™ System minimizes the high cost and complexity of next-generation sequencing and, with Ion Reporter™ Software, facilitates user-defined CNV and aneuploidy detection, with three sensitivity options so that copy number analysis workflows can be tuned to achieve desired levels of sensitivity and specificity.
Mastering RNA-Seq (NGS Data Analysis) - A Critical Approach To Transcriptomic...Elia Brodsky
This workshop will address critical issues related to Transcriptomics data:
Processing raw Next Generation Sequencing (NGS) data:
1. Next Generation Sequencing data preprocessing:
Trimming technical sequences
Removing PCR duplicates
2. RNA-seq based quantification of expression levels:
Conventional pipelines (looking at known transcripts)
Identification of novel isoforms
Analysis of Expression Data Using Machine Learning:
3. Unsupervised analysis of expression data:
Principal Component Analysis
Clustering
4. Supervised analysis:
Differential expression analysis
Classification, gene signature construction
5. Gene set enrichment analysis
The workshop will include hands-on exercises utilizing public domain datasets:
breast cancer cell lines transcriptomic profiles (https://genomebiology.biomedcentral.com/articles/10.1186/gb-2013-14-10-r110),
patient-derived xenograft (PDX) mouse model of tumor and stroma transcriptomic profiles (http://www.oncotarget.com/index.php?journal=oncotarget&page=article&op=view&path[]=8014&path[]=23533), and
processed data from The Cancer Genome Atlas samples (https://cancergenome.nih.gov/).
Team: The workshops are designed by the researchers at the Tauber Bioinformatics Research Center at University of Haifa, Israel in collaboration with academic centers across the US. Technical support for the workshops is provided by the Pine Biotech team. https://edu.t-bio.info/a-critical-approach-to-transcriptomic-data-analysis/
The Main Advantage
The main advantages of flow cytometry over histology and IHC is the possibility to precisely measure the quantities of antigens and the possibility to stain each cell with multiple antibodies-fluorophores, in current laboratories around 10 antibodies can be bound to each cell. This is much less than mass cytometer where up to 40 can be currently measured, but at a higher and slower pace.
Aquatic research
In aquatic systems, flow cytometry is used for the analysis of autofluorescing cells or cells that are fluorescently-labeled with added stains.
This research started in 1981 when Clarice Yentsch used flow cytometry to measure the fluorescence in a red tide producing dinoflagellates
Marine scientists use the sorting ability of flow cytometers to make discrete measurements of cellular activity and diversity, to conduct investigations into the mutualistic relationships between microorganisms that live in close proximity,and to measure biogeochemical rates of multiple processes in the ocean
Cell Proliferation assay
Cell proliferation is the major function in the immune system. Often it is required to analyse the proliferative nature of the cells in order to make some conclusions. One such assay to determine the cell proliferation is the tracking dye carboxyfluorescein diacetate succinimidyl ester (CFSE). It helps to monitor proliferative cells. This assay gives quantitative as well as qualitative data during time-series experiments
Cell counting
Cell sorting
Determining cell characteristics and function
Detecting microorganisms
Biomarker detection
Protein engineering detection
Diagnosis of health disorders such as blood cancers
Flow cytometry can be used for cell cycle analysis to estimate the percentages of a cell population in the different phases of the cell cycle, or it can be used with other reagents to analyze just the S phase.
Why flow cytometry is ideal for cell cycle analysis
Live-cell cycle analysis stains—Vybrant DyeCycle stains
Classic DNA cell cycle stains such as Hoechst 33342 and DRAQ5 for cell cycle analysis, but most of these have limitations that have to be considered when using them in an experiment which is why the Invitrogen Vybrant DyeCycle stains for live-cell cycle analysis were developed.
Fixed-cell cycle analysis stains FxCycle reagents
We offer classic DNA cell cycle stains such as DAPI, PI, and 7-AAD for fixed cell cycle analysis, but these reagents do not cover the full spectrum of laser excitation available.
The FxCycle reagents offer options for the 405 nm (violet) and 633 nm (red) laser thereby increasing the ability to multiplex by freeing up the 488 nm and 633 nm lasers for other cellular analyses such as immunophenotyping, apoptosis analysis, and dead cell discrimination.
Precise—Accurate cell cycle analysis in living cells
Safe—Low cytotoxicity for combining with additional live cell experiments
Cell sort compatible—Easily sort cells based on phase of the cell cycle
the document is about chromosomal analysis technique named array CGH technology, the complete procedure and the result interpretation of chromosomal variation
DNA SEQUENCING METHODS AND STRATEGIES FOR GENOME SEQUENCINGPuneet Kulyana
This presentation will give you a brief idea about the various DNA sequencing methods and various strategies used for genome sequencing and much more vital information related to gene expression and analysis
Aneuploidy in embryos is the leading cause of failure for in vitro fertilization (IVF)
procedures.1 Pre-implantation Genetic Screening (PGS) is used to identify euploid
embryos for implantation to increase successful pregnancies and decrease the number
of cycles required to obtain them. PGS using fluorescence in situ hybridization has
fallen out of favor and been replaced by comparative genome hybridization on
microarrays.1 More recently, high throughput sequencing (HTS) technologies have been
employed for cost-effective PGS on multiple samples.1
We report results using a streamlined Whole Genome Amplification (WGA) and library
generation method followed by HTS and data analysis to detect aneuploidies in single
cells in under 12hr. Using DNA barcodes, we multiplex libraries to reduce the per
sample cost. Our analysis platform compares reads per chromosomal region to an
informatics control built from a baseline of normal cell samples. Using this method, we
show that trisomy of the smallest chromosome (21) can be detected with high sensitivity
and specificity.
Similar to Micro array based comparative genomic hybridisation -Dr Yogesh D (20)
CHAPTER 1 SEMESTER V - ROLE OF PEADIATRIC NURSE.pdfSachin Sharma
Pediatric nurses play a vital role in the health and well-being of children. Their responsibilities are wide-ranging, and their objectives can be categorized into several key areas:
1. Direct Patient Care:
Objective: Provide comprehensive and compassionate care to infants, children, and adolescents in various healthcare settings (hospitals, clinics, etc.).
This includes tasks like:
Monitoring vital signs and physical condition.
Administering medications and treatments.
Performing procedures as directed by doctors.
Assisting with daily living activities (bathing, feeding).
Providing emotional support and pain management.
2. Health Promotion and Education:
Objective: Promote healthy behaviors and educate children, families, and communities about preventive healthcare.
This includes tasks like:
Administering vaccinations.
Providing education on nutrition, hygiene, and development.
Offering breastfeeding and childbirth support.
Counseling families on safety and injury prevention.
3. Collaboration and Advocacy:
Objective: Collaborate effectively with doctors, social workers, therapists, and other healthcare professionals to ensure coordinated care for children.
Objective: Advocate for the rights and best interests of their patients, especially when children cannot speak for themselves.
This includes tasks like:
Communicating effectively with healthcare teams.
Identifying and addressing potential risks to child welfare.
Educating families about their child's condition and treatment options.
4. Professional Development and Research:
Objective: Stay up-to-date on the latest advancements in pediatric healthcare through continuing education and research.
Objective: Contribute to improving the quality of care for children by participating in research initiatives.
This includes tasks like:
Attending workshops and conferences on pediatric nursing.
Participating in clinical trials related to child health.
Implementing evidence-based practices into their daily routines.
By fulfilling these objectives, pediatric nurses play a crucial role in ensuring the optimal health and well-being of children throughout all stages of their development.
India Clinical Trials Market: Industry Size and Growth Trends [2030] Analyzed...Kumar Satyam
According to TechSci Research report, "India Clinical Trials Market- By Region, Competition, Forecast & Opportunities, 2030F," the India Clinical Trials Market was valued at USD 2.05 billion in 2024 and is projected to grow at a compound annual growth rate (CAGR) of 8.64% through 2030. The market is driven by a variety of factors, making India an attractive destination for pharmaceutical companies and researchers. India's vast and diverse patient population, cost-effective operational environment, and a large pool of skilled medical professionals contribute significantly to the market's growth. Additionally, increasing government support in streamlining regulations and the growing prevalence of lifestyle diseases further propel the clinical trials market.
Growing Prevalence of Lifestyle Diseases
The rising incidence of lifestyle diseases such as diabetes, cardiovascular diseases, and cancer is a major trend driving the clinical trials market in India. These conditions necessitate the development and testing of new treatment methods, creating a robust demand for clinical trials. The increasing burden of these diseases highlights the need for innovative therapies and underscores the importance of India as a key player in global clinical research.
Navigating Challenges: Mental Health, Legislation, and the Prison System in B...Guillermo Rivera
This conference will delve into the intricate intersections between mental health, legal frameworks, and the prison system in Bolivia. It aims to provide a comprehensive overview of the current challenges faced by mental health professionals working within the legislative and correctional landscapes. Topics of discussion will include the prevalence and impact of mental health issues among the incarcerated population, the effectiveness of existing mental health policies and legislation, and potential reforms to enhance the mental health support system within prisons.
Health Education on prevention of hypertensionRadhika kulvi
Hypertension is a chronic condition of concern due to its role in the causation of coronary heart diseases. Hypertension is a worldwide epidemic and important risk factor for coronary artery disease, stroke and renal diseases. Blood pressure is the force exerted by the blood against the walls of the blood vessels and is sufficient to maintain tissue perfusion during activity and rest. Hypertension is sustained elevation of BP. In adults, HTN exists when systolic blood pressure is equal to or greater than 140mmHg or diastolic BP is equal to or greater than 90mmHg. The
CHAPTER 1 SEMESTER V PREVENTIVE-PEDIATRICS.pdfSachin Sharma
This content provides an overview of preventive pediatrics. It defines preventive pediatrics as preventing disease and promoting children's physical, mental, and social well-being to achieve positive health. It discusses antenatal, postnatal, and social preventive pediatrics. It also covers various child health programs like immunization, breastfeeding, ICDS, and the roles of organizations like WHO, UNICEF, and nurses in preventive pediatrics.
ICH Guidelines for Pharmacovigilance.pdfNEHA GUPTA
The "ICH Guidelines for Pharmacovigilance" PDF provides a comprehensive overview of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines related to pharmacovigilance. These guidelines aim to ensure that drugs are safe and effective for patients by monitoring and assessing adverse effects, ensuring proper reporting systems, and improving risk management practices. The document is essential for professionals in the pharmaceutical industry, regulatory authorities, and healthcare providers, offering detailed procedures and standards for pharmacovigilance activities to enhance drug safety and protect public health.
Leading the Way in Nephrology: Dr. David Greene's Work with Stem Cells for Ki...Dr. David Greene Arizona
As we watch Dr. Greene's continued efforts and research in Arizona, it's clear that stem cell therapy holds a promising key to unlocking new doors in the treatment of kidney disease. With each study and trial, we step closer to a world where kidney disease is no longer a life sentence but a treatable condition, thanks to pioneers like Dr. David Greene.
Telehealth Psychology Building Trust with Clients.pptxThe Harvest Clinic
Telehealth psychology is a digital approach that offers psychological services and mental health care to clients remotely, using technologies like video conferencing, phone calls, text messaging, and mobile apps for communication.
The Importance of Community Nursing Care.pdfAD Healthcare
NDIS and Community 24/7 Nursing Care is a specific type of support that may be provided under the NDIS for individuals with complex medical needs who require ongoing nursing care in a community setting, such as their home or a supported accommodation facility.
Micro array based comparative genomic hybridisation -Dr Yogesh D
1. Micro-array based Comparative
Genomic Hybridisation(aCGH)
Dr Yogesh D
Resident, Dept. of Anatomy
B J Medical College
Ahmedabad
2. Introduction
• Array Comparative Genomic Hybridisation
(aCGH) is a molecular cytogenetic method for
analysing copy number variations relative to
ploidy level in the DNA of a test sample
compared to a reference sample, without the
need for culturing cells.
3. Principle
• Fluorescent labelled DNA fragments from
patient and control are applied to a microarray
containing tens of thousands of probes fixed
sequentially in triplets, and are allowed to
competitively hybridise with the probes.
• Depending on the copy numbers of the DNA
fragment, varying fluorescence is emitted that
is scanned by an array scanner and an image is
generated of the scan which is analysed by a
software.
4. Method
• The sample collected from the patient is
processed and the DNA is extracted.
• The entire genome is amplified by PCR.
• The amplified genome is fragmented,
denatured and labelled with a fluorescent dye.
5. Method
• Similarly the control DNA fragments are
labelled with a different coloured fluorescent
dye.
• Equal quantities of labelled DNA fragments
from both test and control are then applied on
to the micro array.
• Microarrays contain tens of thousands of
probes that are specifically engineered and
range from 25 base pairs (oligonucleotides) to
200000 base pairs in length.
6. Method
• The DNA fragments from test and control are
allowed to hybridise in PCR machine under
incubation for 36 to 48 hours.
• Excess DNA fragments are washed off.
• The chip is scanned by the chip scanner and an
image of the chip is generated.
• This image is then processed by a software
and a graph is plotted.
7.
8. Reporting Array CGH results
arr[hg19] 16p11.2(29,673,954-30,198,600 )x1
arr -- The analysis was by array-CGH
hg19 -- Human Genome build 19. This is the
reference DNA sequence that the base pair
numbers refer to. As more information
about the human genome is found, new
“builds” of the genome are made and the
base pair numbers may be adjusted.
16p11.2 --- A change was found in band 16p11.2
9. Reporting Array CGH results
(29,673,954-30,198,600 ) -- The first base
pair shown to be missing is number
29,673,954 counting from the left of
the chromosome. The last base pair shown
to be missing is 30,198,600
X1-- means there is one copy of these base pairs,
not two – one on each chromosome 16 – as
you would normally expect.
10. Interpreting Array CGH results
• Array CGH analysis of chromosome 6 for a human
cancer cell line
11. Areas of application
• Prenatal Genetic Diagnosis.
• Cancer cytogenetics.
• Pre-implantation genetic diagnosis.
• Genetic screening of at risk parents.
• Mental retardation of unknown aetiology.
12. Advantages of aCGH
• Can be used to scan the entire genome for
variations.
• Does not require actively multiplying cells.
• Varying probe sizes can be used depending on
the requirement from as short as 25-80bp.
• The resolution offered by aCGH is very high.
• Submicroscopic chromosomal deletions and
duplications can be easily detected.
13. Advantages of aCGH
• Can detect aneuploidies, deletions,
duplications and/or amplifications
simultaneously.
• One assay is equivalent to thousands of
Fluorescent in situ hybridisation experiments.
• Particularly useful in the study of
subtelomeric and pericentromeric
rearrangements.
14. Disadvantages
• Can miss balanced translocations.
• Can miss mosaicism.
• Does not identify tiny deletions or variations.
• Prohibitively expensive.
15. References
• Theisen, A. (2008) Microarray-based comparative genomic hybridization
(aCGH). Nature Education 1(1):45
• Lobo, I. (2008) Chromosome abnormalities and cancer cytogenetics.
Nature Education 1(1):68
• Kannan Thirumulu Ponnuraj (2011). Cytogenetic Techniques in
Diagnosing Genetic Disorders, Advances in the Study of Genetic
Disorders, Dr. Kenji Ikehara (Ed.), ISBN: 978-953-307-305-7, InTech,
• Chial, H. (2008) Cytogenetic methods and disease: Flow cytometry, CGH,
and FISH. Nature Education 1(1):76
• Clancy, S. & Shaw, K. (2008) DNA deletion and duplication and the
associated genetic disorders. Nature Education 1(1):23
• Eichler, E. E. (2008) Copy Number Variation and Human Disease. Nature
Education 1(3):1
Editor's Notes
Array CGH analysis of chromosome 6 for a human cancer cell line. A zoom-in view of an amplified region depicts the distal breakpoint of the copy number gain as well as copy number variants ranging in size from 6,000 to 70,000 base pairs. The top two tracks in the data browser show gene and normal copy number variant annotation. (Source: Peggy Eis, PhD)
aCGH has proven to be a powerful tool for the detection of submicroscopic chromosomal abnormalities in individuals with idiopathic mental retardation and various birth defects. Indeed, several large-scale studies demonstrate that aCGH has a 10%–20% detection rate of chromosomal abnormalities in children with mental retardation/developmental delay with or without congenital anomalies; only 3%–5% of these abnormalities would be detectable by other means. For example, in a study of 8,789 cases analyzed by aCGH, 1,049 (11.9%) had a clinically relevant chromosomal abnormality (Shaffer et al., 2007).
Present on all but the short arms of acrocentric chromosomes 13, 14, 15, 21, and 22, subtelomeric regions have been the subject of a great deal of study because they are relatively gene-rich (Saccone et al., 1992) and are prone to rearrangement by a number of mechanisms (Ballif et al., 2003, 2004). Moreover, rearrangement of subtelomeric regions has been suggested to represent a high proportion of abnormalities in individuals with idiopathic mental retardation.
The second class of CNVs includes relatively rare variants that are much longer than CNPs, ranging in size from hundreds of thousands of base pairs to over 1 million base pairs in length. Also know n as microdeletions and microduplications, these variants usually have a much more recent origin w ithin a family. These CNVs may have arisen during production of the sperm or egg that gave rise to a particular individual, or they may have been passed dow n for only a few generations w ithin a family. These large and rare structural variants have been observed disproportionately in patients w ith mental retardation, developmental delay, schizophrenia, and autism (de Vries et al., 2005, Sharp et al. 2006, Sebat et al., 2007, Walsh et al., 2008). Their appearance in such patients has led to speculation that large and rare CNVs may be more important in neurocognitive diseases than other forms of inherited mutations, including single nucleotide substitutions.