scholarly journals Identification of Genetic Markers Using Polymerase Chain Reaction (PCR) in Graves' Hyperthyroidism

10.5772/37491 ◽  
2012 ◽  
Author(s):  
P. Veeramuthumari ◽  
W. Isabel
2019 ◽  
Vol 4 ◽  
pp. 25-34
Author(s):  
Oleksandr Glavatskyi ◽  
Irina Vasileva ◽  
Olena Galanta ◽  
Hennadii Khmelnytskyi ◽  
Irina Shuba ◽  
...  

Intracerebral malignant brain tumors remain one of the most complex problems of neuro-oncology. Today, promising results of the use of targeted drugs have been received, which determine the important diagnostic and predictive value of molecular genetic markers of glial and metastatic brain tumors. Aim: The study of the prevalence of MGMT (O6-methylguanine-DNA methyltransferase) and PTEN (phosphatase and tensin homologue deleted on chromosome 10) gene expression by real time polymerase chain reaction in tumor tissue of gliomas and brain metastases. Materials and methods: From thirty patients were received tumor material (29 cases of glioma III-IV degree of anaplasia and one case of metastatic brain lesion of adenocarcinoma). The normalized expression of MGMT and PTEN genes was determined by real-time polymerase chain reaction. Results: In all 30 (100 %) patients with tumor fragments, we determined normalized expression of MGMT and PTEN genes. In most cases, 53 % of the observations (16 out of 30 patients) showed a low normalized expression of MGMT gene (<40 c. u.) and a low normalized PTEN expression rate of 73 % (22 out of 30 patients) (<40 c. u.). The average expression level of the MGMT gene in the range from 40 to 100 c. u. (6/20 % of patients) was considered prognostic favourable for the response to temozolomide chemotherapy. Conclusions: The study of MGMT gene expression, a chemotherapy marker for temozolomide, indicates a trend toward correlation between expression levels and therapeutic efficacy. The study of the expression of the PTEN gene, the blocker of the PI3K / AKT signal pathway, indicates a different degree of expression of this enzyme in the tumour samples studied. The predictive value of the indicator for target therapy is appropriate in comparison with the EGFR mutation. Further profound analysis of the results is required with increasing number of sampling and observation period.


Author(s):  
Surya Kannan ◽  
◽  
Johan Ericsson ◽  
Nazariy Souchelnytskyi ◽  
Serhiy Souchelnytskyi ◽  
...  

Introduction. Polymerase chain reaction (PCR)-based diagnostic tests use purifi ed nucleic acids (NAs) from clinical samples. The NAs purifi cation step adds time, cost, and aff ects the quality of testing. The objective of this study was to develop a protocol for direct use of saliva in tests for genetic markers, without purifi cation of nucleic acids. Methods. PCR, real-time RT-PCR and isothermal amplifi cation tests were used for direct detection of genetic markers, without purifi cation of nucleic acids. Results. We report a protocol for the direct detection of genetic markers in saliva. The protocol is based on a collection of saliva in a solution containing a detergent and ethanol and is compatible with isothermal amplifi cation (LAMP), real-time RT-PCR and RT-PCR. SARS-CoV-2 and GAPDH markers were used as reference markers. We observed that mild detergents allow effi cient detection of external reference and intracellular endogenous markers, while strong detergent, e.g. sodium dodecyl sulfate, inhibited the PCR reaction. Under these conditions, saliva samples can be stored for 24 h at +4°C or –18°C with the preservation of markers. Storage at room temperature led to the deterioration of marker detection. Snap heating of saliva samples at the time of collection, followed by storage at room temperature, provided partial protection. Conclusion. The protocol presented in this report describes the collection and storage of saliva for direct detection of genetic markers and is compatible with PCR and LAMP tests.


1991 ◽  
Vol 63 (1) ◽  
pp. 2-15 ◽  
Author(s):  
Rebecca. Reynolds ◽  
George. Sensabaugh ◽  
Edward. Blake

1996 ◽  
Vol 87 (7) ◽  
pp. 669-675 ◽  
Author(s):  
Federico Canzian ◽  
Toshikazu Ushijima ◽  
Minoru Toyota ◽  
Yoko Hosoya ◽  
Takashi Sugimura ◽  
...  

Genome ◽  
1994 ◽  
Vol 37 (2) ◽  
pp. 244-248 ◽  
Author(s):  
David S. Haymer ◽  
Donald O. McInnis

We have used the polymerase chain reaction (PCR) and the random amplified polymorphic DNA (RAPD) method to identify DNA polymorphisms that can be used as genetic markers to characterize populations of the Mediterranean fruit fly, Ceratitis capitata. In this study, RAPD markers have been used to resolve genetic variability between populations of this major agricultural pest species. The populations analyzed represent either laboratory stocks or wild collections originating from different geographic localities. Using the same set of individual flies from each of several populations, we show that the use of different primers in the RAPD method permits detection of different levels of population differentiation. We show results from RAPD primers (e.g., primer 14) that identify regions of the genome (through PCR amplification) that are essentially monomorphic in all flies originating from a particular geographic locality. We also show RAPD primers (e.g., primer 67) that identify what appear to be highly variable regions of the genome. We have used primers of this type to produce genetic markers that can distinguish even between laboratory versus wild populations as well as subpopulations of flies from more broadly defined geographic localities, such as within the Hawaiian islands. These results show that the RAPD method is a broadly applicable, high resolution method for documenting genetic variability within and between populations of insect pest species.Key words: RAPD–PCR, genetic markers, Ceratitis capitata, Mediterranean fruit fly populations.


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