The association between the presence of a rare
BRCA2 mutation and the possibility of developing
endometrial cancer due its inheritance in a
Romanian population and the utilization of saliva
samples to detect the mutation.
Name
ID:
Tutorial group 00
Faculty of Health, Medicine and Life Sciences (FHML)
Maastricht University
Course BBS1005
Practical training session: 12th May 2022
16th June 2022
1
, 1. Introduction
For decades it has been a well-known fact that the prolonged accumulation of
changes in the DNA sequence of animal cells, otherwise known as mutations, can
result into cancers (1). Apart from spontaneous mutations, family history has been
found to play a role in carcinogenesis (2). Numerous methods for extracting genomic
DNA and analyzing it for mutations have been developed, however this study
focuses primarily on DNA obtained via saliva samples and followed by polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis for
carcinogenic mutations. Specifically in the breast cancer 2 genes (BRCA2) in a
Romanian population. A tendency to proliferate uncontrollably and an invasive
behavior, which causes them to metastasize to different organs are common
characteristics among cancer cells (1). Point mutations in specific genes of somatic
cells have been identified that lead to cells with such characteristics (3). In order to
detect these mutations methods such as PCR-RFLP followed by gel electrophoresis
can be used. Nonetheles, a genomic DNA sample must be first isolated from the
individual testing for a certain hereditary mutation, which will have to meet specific
purity and concentration standards in order for the PCR-RFLP technique to be
carried out. Peripheral blood has been the ideal source of genomic DNA in genetic
studies and genetic testing, however due the invasive nature of this technique and
the possible risks concerning the people tested, an alternative technique based on
buccal cells has been developed (4, 5). In this technique, genomic DNA is isolated
from buccal cells, which are found in the inner side of the cheeks in the oral cavity
and are present in a person’s saliva sample (4). It is a cost effective, non-invasive
method, which has been found to yield sufficient DNA quality and quantity for
research and clinical testing (4). In a study directed by Küchler E.C. and colleagues,
it was concluded that DNA isolated from buccal cells in saliva provided quality DNA
samples of sufficient amount for caring out successful PCR-RFLP reactions (4).
Küchler E.C. and colleagues also reported that DNA quality may be prone protein
contamination when using this DNA isolation method (4). In order to further
investigate and ensure whether DNA isolated from saliva samples can be of
acceptable purity and quantity for PCR-RFLP reactions, like in the case of Küchler
E.C. and colleagues, Maastricht University of the Netherlands decided to direct its
own DNA isolation research using samples from a cohort of first year Biomedical
Sciences students.
Acquiring enough high-quality genomic DNA is a perquisite for a successful PCR
amplification and RFLP analysis thereafter (4). The PCR-RFLP method followed by
2
BRCA2 mutation and the possibility of developing
endometrial cancer due its inheritance in a
Romanian population and the utilization of saliva
samples to detect the mutation.
Name
ID:
Tutorial group 00
Faculty of Health, Medicine and Life Sciences (FHML)
Maastricht University
Course BBS1005
Practical training session: 12th May 2022
16th June 2022
1
, 1. Introduction
For decades it has been a well-known fact that the prolonged accumulation of
changes in the DNA sequence of animal cells, otherwise known as mutations, can
result into cancers (1). Apart from spontaneous mutations, family history has been
found to play a role in carcinogenesis (2). Numerous methods for extracting genomic
DNA and analyzing it for mutations have been developed, however this study
focuses primarily on DNA obtained via saliva samples and followed by polymerase
chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis for
carcinogenic mutations. Specifically in the breast cancer 2 genes (BRCA2) in a
Romanian population. A tendency to proliferate uncontrollably and an invasive
behavior, which causes them to metastasize to different organs are common
characteristics among cancer cells (1). Point mutations in specific genes of somatic
cells have been identified that lead to cells with such characteristics (3). In order to
detect these mutations methods such as PCR-RFLP followed by gel electrophoresis
can be used. Nonetheles, a genomic DNA sample must be first isolated from the
individual testing for a certain hereditary mutation, which will have to meet specific
purity and concentration standards in order for the PCR-RFLP technique to be
carried out. Peripheral blood has been the ideal source of genomic DNA in genetic
studies and genetic testing, however due the invasive nature of this technique and
the possible risks concerning the people tested, an alternative technique based on
buccal cells has been developed (4, 5). In this technique, genomic DNA is isolated
from buccal cells, which are found in the inner side of the cheeks in the oral cavity
and are present in a person’s saliva sample (4). It is a cost effective, non-invasive
method, which has been found to yield sufficient DNA quality and quantity for
research and clinical testing (4). In a study directed by Küchler E.C. and colleagues,
it was concluded that DNA isolated from buccal cells in saliva provided quality DNA
samples of sufficient amount for caring out successful PCR-RFLP reactions (4).
Küchler E.C. and colleagues also reported that DNA quality may be prone protein
contamination when using this DNA isolation method (4). In order to further
investigate and ensure whether DNA isolated from saliva samples can be of
acceptable purity and quantity for PCR-RFLP reactions, like in the case of Küchler
E.C. and colleagues, Maastricht University of the Netherlands decided to direct its
own DNA isolation research using samples from a cohort of first year Biomedical
Sciences students.
Acquiring enough high-quality genomic DNA is a perquisite for a successful PCR
amplification and RFLP analysis thereafter (4). The PCR-RFLP method followed by
2
