To install StudyMoose App tap and then “Add to Home Screen”
Save to my list
Remove from my list
This laboratory report outlines the methodology for detecting T.N.F-α (238 G/A) gene polymorphism using TaqMan probe SNP assay and real-time PCR. The study also includes the assessment of cardiovascular liability index, anthropometric characteristics, and medical characteristics among the subjects. The analysis was conducted on 50 psoriatic subjects and 50 controls. The results provide insights into the genetic variations associated with psoriasis and its potential correlation with cardiovascular risk factors.
Psoriasis is a chronic autoimmune skin disease that has been associated with various genetic factors.
T.N.F-α (238 G/A) gene polymorphism is one such factor of interest. This report presents the methodology for its detection and explores the relationship between genetic variations and cardiovascular risk factors.
Genomic DNA was extracted from stored samples using DNA purification kits following the manufacturer's protocol. The following steps were involved:
| Step | Description |
|---|---|
| 1 | 200 μl pure ethanol added to the sample, followed by pulse-vortex for 15 seconds |
| 2 | Centrifugation to remove drops from inside the lid |
| 3 | Application of the mixture to QIAamp Spin Column without wetting the rim |
| 4 | Centrifugation at 6000 xg for 1 minute |
| 5 | Buffering factors A.W.1 and A.W.2 added sequentially |
| 6 | Centrifugation at full speed (20,000 xg) for 3 minutes |
| 7 | Buffering factor A.E added and incubation at room temperature for 1 minute |
| 8 | Centrifugation at 6000 xg for 1 minute to collect the DNA filtrate |
The assay is based on the TaqMan probe SNP assay method.
Verified writer
Proficient in: Biology
4.7 (348)
“ Amazing as always, gave her a week to finish a big assignment and came through way ahead of time. ”
It utilizes a probe containing a fluorescent editor dye and a quencher dye to detect genetic variations.
The following steps outline the process:
The TaqMan probe contains a fluorescent editor dye at the 5' end and a quencher dye at the 3' end. When the probe is intact, fluorescence is inhibited. During PCR, the probe is cleaved, separating the dyes, allowing detection of edited dye fluorescence (F.A.M. for the T allele and V.I.C. for the G allele).
An oligonucleotide probe is used with edited fluorescent and quencher dyes. The probe is cleaved during PCR, allowing detection of fluorescence.
The probe is cleaved, releasing the edited dye, leading to increased fluorescence for G and A alleles of the T.N.F-α gene.
The amplified products are analyzed using TaqMan probes for G and A alleles, with fluorescence intensities used for detection.
The cardiovascular liability index was determined using various parameters:
The PASI score was calculated to evaluate the severity of psoriasis. It considers the extent of erythema, thickness, and scaling at four anatomic sites and assigns numerical values to these parameters.
PASI = (0.1 * (Eh + Ih + Dh) * Ah) + (0.2 * (Eu + Iu + Du) * Au) + (0.3 * (Et + It + Dt) * At) + (0.4 * (El + Il + Dl) * Al)
Where E = erythema, I = infiltration, D = desquamation, A = area, h = head, t = trunk, u = upper extremities, and l = lower extremities.
The DLQI was used to assess the impact of psoriasis on the quality of life. It involves a scoring system based on the patient's responses to specific questions.
Data were analyzed using SPSS software. Descriptive statistics, t-tests, chi-square tests, and Fisher exact tests were used for data analysis.
The study included 50 psoriatic subjects and 50 controls. Key findings are summarized below:
The mean age of psoriatic subjects was 39.75 years, with a male-female ratio of 52% to 48%. Family history of psoriasis was present in 12% of cases.
| Characteristic | Mean ± SD | Minimum | Maximum |
|---|---|---|---|
| Age | 39.75 ± 14.20 | 18.00 | 66.00 |
| Sex (Male) | 52.0% | - | - |
| Family History (Yes) | 12.0% | - | - |
Anthropometric characteristics of the psoriatic subjects were as follows:
| Characteristic | Mean ± SD | Minimum | Maximum |
|---|---|---|---|
| Weight (kg) | 82.36 ± 17.80 | 55.00 | 120.00 |
| Height (cm) | 165.34 ± 8.94 | 147.00 | 192.00 |
| BMI | 30.20 ± 6.84 | 20.76 | 54.78 |
| Waist Circumference (cm) | 100.50 ± 16.30 | 62.00 | 132.00 |
Medical characteristics among psoriatic subjects included carotid duplex measurements, blood pressure, hypertension, diabetes, and ECG results.
| Characteristic | Mean ± SD | Minimum | Maximum |
|---|---|---|---|
| Carotid Duplex | 0.09 ± 0.02 | 0.07 | 0.15 |
| Systolic Blood Pressure | 120.71 ± 13.42 | 90.00 | 155.00 |
| Diastolic Blood Pressure | 78.67 ± 10.19 | 50.00 | 95.00 |
| Hypertension (Yes) | 10.0% | - | - |
| Diabetes (Yes) | 8.0% | - | - |
| ECG (Abnormal) | 20.0% | - | - |
The results of this study indicate a potential association between T.N.F-α (238 G/A) gene polymorphism and psoriasis. Furthermore, cardiovascular risk factors, including BMI, waist circumference, and blood pressure, were observed among psoriatic subjects. These findings suggest the importance of monitoring cardiovascular health in psoriasis patients.
This laboratory report highlights the methodology for detecting T.N.F-α (238 G/A) gene polymorphism in psoriatic subjects and the assessment of cardiovascular liability index. The study provides valuable insights into the genetic and cardiovascular factors associated with psoriasis, contributing to a better understanding of the disease.
Detection of T.N.F-α (238 G/A) Polymorphism by Real-time PCR. (2024, Jan 22). Retrieved from https://studymoose.com/document/detection-of-t-n-f-238-g-a-polymorphism-by-real-time-pcr
👋 Hi! I’m your smart assistant Amy!
Don’t know where to start? Type your requirements and I’ll connect you to an academic expert within 3 minutes.
get help with your assignment
