Application of PCR Techniques for Detecting Transcripts in Leukaemia

This report delves into the application of Polymerase Chain Reaction (PCR) techniques for detecting transcripts associated with translocated genes in leukaemia, a blood cancer characterized by the abnormal proliferation of white blood cells. The document provides an overview of leukaemia, including its symptoms like fatigue, bleeding, and infections, as well as diagnostic methods such as blood tests, staging, and imaging. It explores PCR's role in identifying specific gene areas, detailing the steps involved, from DNA isolation and primer design to enzyme selection and amplicon analysis. The report also covers treatment options like chemotherapy, radiation therapy, and stem cell transplantation. Risk factors, prevention strategies, and the importance of early diagnosis are also discussed. This comprehensive analysis aims to provide a clear understanding of how PCR is used in the diagnosis and management of leukaemia.

PCR techniques for the detection
of transcripts on translocated
genes in leukaemia

Table of Contents
INTRODUCTION...........................................................................................................................3
MAIN BODY..................................................................................................................................3
Leukaemia diseases and PCR techniques. .............................................................................3
Symptoms of Leukaemia:.......................................................................................................5
Diagnostic method used in Leukaemia:.................................................................................5
Risk factors: ...........................................................................................................................7
Prevention from leukaemia diseases: ....................................................................................9
PCR techniques for the detection of transcripts on translocated genes in leukaemia............9
CONCLUSION ............................................................................................................................10
REFERENCES..............................................................................................................................11

INTRODUCTION
Leukaemia is a blood cancer which is develop in white blood cell. The white blood
cell are increasing their growth and enhance their number in blood then it develop Leukaemia.
As per this, red blood cell and platelets must be gain proper oxygen for their functioning, and
they need to be healthy. In this, the extra blood cell lead to follow abnormal growth, so on the
basis of this, they form cancer cell (Bhat and Rao 2020). They rise with various symptom and
formulate various complication in body. In this, the polymerase chain reaction is method which
is used in the detection of specific gene areas, this detection is based on various step and
confirmatory stages. In this report, the use of polymerase chain reaction in leukaemia is analyse
and also some of the prevention techniques which is must followed in order to procure the
abnormal growth of cell in blood. The associated symptoms and risk factor are also
demonstrated in this report.
MAIN BODY
Leukaemia diseases and PCR techniques.
Leukaemia is a blood cancer which refers to increasing white blood cell. Every healthy
person required red blood cell and platelets in body. In order to this, for expanding their life span
them this type of disease in must prevented which create issues, formulate healthy life habits
adopted along with the healthy food. Variation in the blood plasma contain cause this problem
due to the higher production of WBC and less production of RBC. For individuals, it is essential
to take healthy and iron rich food and this habit help to individual in increase of RBC in blood
plasma. In blood, there are various essential substance which work as to protect body from
various pathogens and helps in several organ functioning, but inappropriate composition of the
blood plasma can cause problems such as leukaemia which is start in the blood by increasing the
formation rate as per the abnormal growth, it cause the over production of white blood cells
(Boyd and et. al., 2017).
White blood cell are generated and as with this there are number of cell which is in
lymph nodes, spleen, thymus gland. This disease majorly divided in two categories such as
myeloid cells cancer which refers to myelogenous leukaemia and another one is lymphocytic
leukaemia. There are different various of techniques which usually used in the cancer treatment
and diagnosis such as PCR techniques and many more. PCR known as Polymerase Chain

Reaction, this is the diagnostic techniques or molecular techniques which helps in the detection
of leukaemia. It is also said to be reverse transcription PCR and Reverse transcription
quantitative real time PCR. It is a techniques which are used in health care for detection and
diagnosis of leukaemia diseases. It aid to check the DNA which result as specific identification
of this disease in medial history of patient such as hereditary and genetic disorder. In this
techniques, small amount of sample is to be taken and analyse through this method which
provide result after a short period of time. As per this test and diagnosis the result is visible after
few days. It is useful technique for detection of leukaemia from the body which refers to
identification of genes in order to know about particular aspect of diseases.
The polymerase chain reaction is a sensitive and efficient method for targeting a single
DNA from million of copies (Chan and et. al., 2017). In addition, the target DNA detection and
amplification which is used by Polymerase chain reaction is an major step fro cloning, finding
gene expression and many more. Polymerase chain reaction has various application in several
field such as in analysis of cancer, biotechnology and also play active role in research processes.
In this they usually detect cancer cell by following five step which is mentioned below:
Step1 DNA isolation: DNA isolation is first step in technique of PCR, whereas in this the
genomic DNA is isolated or bacteria plasmid. In overall, now the selection of nucleic acid
purification of product vis taken in order to provide maximum yield, purity and integrity from
sample which is collected.
Step2 Primer design: In this the good sequence is chosen and high quality of primer for PCR
reaction. The length of 18-30 nucleotide for primer is best for this. As per this, the melting
temperature of primer should be between 65 degree Celsius and 75 degree Celsius and this will
up and down with 10 percent of melting temperature.
Step3 Enzyme selection: DNA polymerase is an important element of PCR. It play a vital role in
synthesizing DNA strand. So in this context, it is compulsory to choose high quality of enzyme
for produce good results.
Step4 Thermal cycling: In this PCR consisting of warming and chilling cycles for DNA
amplification. As per this, the success and failure of this techniques is totally depend on
choosing the right thermal cycler. Some of the cases automation of process is taken to complete
this test.

Step5 Amplicon analysis: As per this, nucleic acid electrophoresis is a simple approach is taken
to separate the PCR workflow. In the electrophoresis selecting an right nucleic acid can improve
and enhance the result and access more application to drawn.
Symptoms of Leukaemia:
There are several symptoms of this diseases which are faced by patient in condition of
blood cancer. Anaemia is diseases so similar types of symptoms such as weakness and fatigue in
body. It considers as patient feel pallor and general feeling of illness (Erwanto and et. al., 2018).
There are numbers of factors which are discussed below:
 The symptoms in this is bleeding from gums and nose.
 Individual feel or faced infection in order to sore throat or bronchial infection, headache,
low-fever, mouth sores and skin rash.
 It considered as swollen lymph node which are impact on neck, armpits and groin.
 Suspected person face issues and problems such as loss of weight and loss of appetite.
 Patient feel such as swelling in lower ribs and feel discomfort.
 The major impact on white blood cell. So, as per this, the increasing of white blood cell
as compare than red blood cell. In this, patient also faced retinal haemorrhage, ringing of
the ears, mental instability, stroke and many others.
 It considered as individual feel extensive weakness in body and many other complication
in body.
It is essential for person who faced these type of symptoms then they must concern with
doctor their complication and in this context person must take confirmatory test and diagnostic
tool. Leukaemia is also spread in multiple organs of body because they have tendency to
metastasis their cell in various zone on body. Leukaemia is a blood cell cancer and their major
complication is related with central nervous system. Accordance to this, patient faces numbers of
problems such as headache, nausea and vomiting, confusion, pain in muscles and seizures. It
frequently spread in another body parts such as lungs, gastrointestinal tract, heart, kidneys and in
many part where they possibly reached.
Diagnostic method used in Leukaemia:
It is necessary for those individual in order to know about the risk factor and symptoms
of this diseases. So, accordingly make diagnose as in result to improving health. It is consist of

several test and diagnostic method which are used in this diseases as well as beneficial for
patient which is discussed below.
Tests: It includes several tests which are recommended by doctor in order to physical
examination which are necessary those who is facing symptoms regards with leukaemia. It
involves tests such as cells test which works as to analyse red blood cell, white blood cell and
platelets (Green and Sambrook 2019). In addition to this, individual feel discomfort in lymph
nodes as well as body pain then they must refer for the leukaemia based test. On that basis test
are necessary for suspected person which refers to take small sample of blood and investigate in
order to confirmatory test for leukaemia.
Staging: In pre stage diagnostic leukaemia doctors moved to second stage which refers to
staging. It involves to know about the cancer cell look like so used as microscope techniques in
order to different kinds of cell involved. On that stage the count of WBC in time diagnosis as
compare to RBC. It involves various techniques in this stage in such a way to count cell such as
AML and CML.
Assessing progression: It considered as several numbers of tests in order to examine this
diseases. It includes they are affecting numbers of multi organ which result as cytometry
examination which refers to it is kind of tool and techniques to test DNA. It is test effectively
used in order to determining growth rate of cell as well as liver test. Because leukaemia cell are
spread and infect liver. Lumbar puncture is kind of tool which are used by lab attended in order
to inserting thin needle in lower back to test this. This is a technique which is useful because it
help to know about spreading aspect of cancer or not. It considered as imaging tests such as X-
rays, ultrasounds and CT scans in order to know about effect on other body part or not, which is
caused by leukaemia.
Treating leukaemia: This is stage where suspected person confirmed for leukaemia. So, as per
their confirmation the treatment of this diseases is started. This disease have proper treatment so
on the basis of this they depends on various stages because cancer has several stages. On that
basis doctors provides treatment of patient such as leukaemia in many forms. It involves various
forms of this diseases. So, they grow slowly accordingly patient follow the prescription of doctor
as well as there is not need of immediate treatment. There are various tools and techniques which
are used by doctors in leukaemia diseases which are mentioned below.

Chemotherapy: It is kind of drugs treatment and medicine which are used by doctor for
cancer patient. In order to kill the fast growing leukaemia cells. Majorly doctors study and
analyse the type of leukaemia as well as type of diseases. According to that they decide the
combination of drugs for chemotherapy .
Radiation therapy: It includes doctors recommended this therapy for cancer patient because it is
helpful to reducing or damage leukaemia cell in from body. It is a type of beam radiation therapy
which use to target and kill the abnormal cells but it can be harmful for body in some case.
Radiation have an ability to degrade the DNA and cause mutation, so this method is use in very
fewer cases .
Steam cell transplantation: It is technique and tool which known as bone morrow
transplantation in order to replaced damage bone morrow with healthy bone marrow.
Biological or immune therapy: It is another stage of treatment which refers to providing
medicine in order to boost up immunity system as well as attack on cancer cells. In this disease
doctor giving suggestion to patient for the use of targeted therapy such as uses medication
because it is good for cancer patient.
Therefore, it includes various treatment which are useful and beneficial for leukaemia
patient, doctors provide effective treatment for cancer patient. It involves various techniques
which are implemented by doctor and physician to take proper care of cancer patient as well as
delivering best services for patient. This kinds of diseases majorly depends on stages. Because
cancer is divided into numbers of stages (Irani and et. al., 2020). So, accordingly that various
techniques are used by doctor in order to tested or diagnose this diseases. For increasing chances
of recovery from leukaemia diseases are taken when right methods of treatment should be
selected as per the analysis which is depend on condition of patient.
Risk factors:
There are various risk which enhance chance of leukaemia and blood cancer such as old
age, past history of blood disorders and chromosome mutation and many other reason. There are
several reason which are including as risk factor and greater chances of leukaemia which are
mentioned below.
Previous cancer treatment: It considered as leukaemia diseases have more chances to occur due
to the medical history of cancer treatment, so past history analysis of patient related to the cancer
treatment is important, if patient has taken the any kind of treatment such as radio therapy or

chemotherapy. It considered as majorly doctors has to focus on past history to know about the
diseases .
Genetic disorders: Genetic mutation or disorder is can be related to the cause of leukaemia
because, mutation in cell can change their growth method and cause abnormal growth. So, this
factor can be consider in the process of diagnosis (Lagunas-Rangel and et. al., 2017).
Exposures to certain chemicals: Chemical component can be a causes of leukaemia because
some of the chemical can cause mutation of cells and then mutation result as the abnormal cell
growth.
Smoking: This can be another cancer causing factor which posses 30 % chances of leukaemia
due to presence of nicotine.
Family history: This is important factor for suspected person, it is essential to check their family
history because several cancer can be heredity. Doctor may first consider patient and then their
family medical history along with the medical test in order to identify the causes of this
abnormality.
There are several advanced developments and techniques which are recently developing
in order to diagnostic leukaemia. It involves various tools and techniques which are utilised by
physician and doctor to get more advancement in diagnostic test which are beneficial for cancer
patient. It provide more chance to patient recovery. It considered as various development which
are helpful for treatment in cancer patient due to doctors. It consist new therapies approved by
National Cancer Institute to get more chance in patient recovery. It includes various therapy
which is help to reduce the risk factor of cancer.
It includes various drugs, from these two drug which are prescribed in order to used as
treatment of patient such as IDH2 protein and liposome cytarabine daunorubics CPX-351 which
known as Vyxeos. These drug are used as treatment of leukaemia patient as well as in
combination with chemotherapy. The drug such as Vyxeos which are more effcetive and have
higher therapeutic activity as compared with other drug because it helps to minimizing the risk
of cancer. This kind of drug which are approved from Food and Drug Administration as well as
approved from companion diagnostic test (Shlush and et. al., 2017). This all drug are more
effective because it play beneficial role in cancer patient in order to treatment with proper
medicine and drug. It is more effective dose for this drug. So, the reaction on patient body such
as providing treatment help to haematological recovery. It is process which refers that no sign in

body count the blood cells in order to RBC, WBC and platelets. It is long process because it will
take 56 day of time. There are various common side effects such as jaundice, nausea, fever as
well as impact on respiratory system. This kind of treatment which shows the best result as well
as most of patient recovery (Zhang and et. al., 2020).
Hence, it includes various advancement in drug and diagnostic test and which are
approved by National Cancer Institute. This treatment which are adapting effectively in order to
providing best treatment for cancer patients as well as using new method effectively and
efficiently.
Prevention from leukaemia diseases:
There are several preventions techniques are necessary for leukaemia which is prescribed
by doctor and it is mandatory by patient to follow this medication as per given instruction. Here,
some of the preventions technique which are mentioned and its is necessary for each and every
individuals.
Be a non-smoker: The effect which is usually caused and promote the chance of cancer is
smoking. So as per this, every person must focus on their proper health and this reduce chances
of cancer. Avoid smoking in respect with healthy life.
Maintain a health: It involves most of physician advice to maintain health and body weight as
per the body mass index. Because unhealthy diet and obesity can increase the risk of this
diseases. It is essential to eat healthy food and also preparing appropriate diet plan.
In addition to this, the above discussed techniques is useful to reduce the risk of cancer.
PCR techniques for the detection of transcripts on translocated genes in leukaemia
As per the recurrent chromosomal translocation have some proper proportion of myeloid
and lymphoid leukaemia, in aspect to this, the formation of unique gene are happen. The
determination of various fusion gene which is abnormal in nature have their diagnostic and
prognostic importance (Felt and et. al., 2017). Moreover, identification of translocation fusion
gene abnormalities is just a common factor by reverse transcription polymerase chain reaction
amplification encompasses the junction which have specific areas for the chimeric mRNA
matter. As per this, the interpretation of electrophoretic band which have visualise pattern and
have ability to production of non specific PCR amplicons can frequently synthesizing an
difficulties in their analysis. To eradicate this problem there are number of labs which work to
add blot hybridization step by using proper obignonucleotide probe with isotopic or non-isotopic

nylon membrane for the detection. As per this, the specific blot and probe method increase the
effectivity. This is more costly and time consuming. In such a way the specific probe
hybridization detection is target on PCR amplicons and furthermore. By optimizing various
aspect in both the PCR and post PCR analysis (Agarwal and et. al., 2020).
CONCLUSION
From the above report, it has been concluded that leukaemia is diseases cause at the site
blood tissues due to the production of abnormal white blood cell and reduction of the red blood
cells . For the analysis and identification disease in patient PCR technique is which is consider as
the polymerase chain reaction method it aid to study the DNA replication of the cell, through
which growth rate of the cells are identify and then according to that result is consider. At
present developing technology there is the huge efforts made on development of cancer
treatment techniques such as identically targeted KO-539 menin inhibitor use for myeloid
leukaemia treatment.

REFERENCES
Books and Journal
Bhat, A.I. and Rao, G.P., 2020. Polymerase chain reaction. In Characterization of Plant
Viruses (pp. 323-345). Humana, New York, NY.
Boyd and et. al., 2017. Acute myeloid leukaemia disrupts endogenous myelo-erythropoiesis by
compromising the adipocyte bone marrow niche. Nature cell biology, 19(11), pp.1336-
1347.
Chan and et. al., 2017. Disseminated fusarium infection after ibrutinib therapy in chronic
lymphocytic leukaemia. Annals of hematology, 96(5), pp.871-872.
Erwanto and et. al., 2018. Identification of pig DNA in food products using polymerase chain
reaction (PCR) for halal authentication-a review. International Food Research
Journal, 25(4), pp.1322-1331.
Green, M.R. and Sambrook, J., 2019. Polymerase chain reaction. Cold Spring Harbor
Protocols, 2019(6), pp.pdb-top095109.
Irani and et. al., 2020. Successful treatment‐free remission in chronic myeloid leukaemia and its
association with reduced immune suppressors and increased natural killer cells. British
journal of haematology, 191(3), pp.433-441.
Lagunas-Rangel and et. al., 2017. FLT3–ITD and its current role in acute myeloid
leukaemia. Medical oncology, 34(6), p.114.
Shlush and et. al., 2017. Tracing the origins of relapse in acute myeloid leukaemia to stem
cells. Nature, 547(7661), pp.104-108.
Agarwal and et. al., 2020. Clinicopathological, microbiological and polymerase chain reaction
study in a case of Nocardia scleritis. Ocular Immunology and Inflammation, pp.1-5.
Felt and et. al., 2017. The utility of real-time quantitative polymerase chain reaction genotype
detection in the diagnosis of urinary tract infections in children. Clinical
pediatrics, 56(10), pp.912-919.
Zhang and et. al., 2020. Universal linker Polymerase Chain Reaction-triggered Strand
Displacement Amplification visual biosensor for ultra-sensitive detection of
Salmonella. Talanta, 222, p.121575.