scholarly journals Upregulation of the miRNA-155, miRNA-210, and miRNA-20b in psoriasis patients and their relation to IL-17

2020 ◽  
Vol 34 ◽  
pp. 205873842093374 ◽  
Author(s):  
Mohamed El-Komy ◽  
Iman Amin ◽  
Marwa Safwat El-Hawary ◽  
Dina Saadi ◽  
Olfat Shaker
Keyword(s):  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Epigenetic Mechanisms ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Lesional Skin ◽  
Immune Mediated ◽  
Elisa Technique ◽  
Polymerase Chain ◽  
The Relationship

Psoriasis is an immune-mediated disease, with genetic background and triggering environmental factors; however, several gaps are still present in understanding the intertwined relationship between these elements. Epigenetic mechanisms, including microRNAs (miRNAs), play an important role in the pathogenesis of psoriasis. The relationship between interleukin (IL)-17, a key cytokine in psoriasis, and these epigenetic mechanisms still needs to be elucidated. This study aimed at assessing the expression of miRNA-155, miRNA-210, and miRNA-20b in skin and sera of psoriasis patients in relation to IL-17 levels. For 20 psoriasis patients and 20 matching controls, the expression of miRNA-155, miRNA-210, and miRNA-20b was assessed using real-time polymerase chain reaction (RT-PCR), whereas IL-17/IL-17A levels were measured using quantitative enzyme-linked immunosorbent assay (ELISA) technique. MiRNA-155 expression was significantly higher in lesional skin compared to controls ( P = 0.001). MiRNA-210 expression was significantly higher in both, lesional skin ( P = 0.010) and sera of patients ( P = 0.001) in comparison with controls. A statistically significant positive correlation was found between serum miRNA-210 expression and serum levels of IL-17/IL-17A ( P = 0.010, rs = 0.562). MiRNA-20b lesional and non-lesional expression was significantly higher than controls ( P < 0.001; P = 0.018). In conclusion, the expression of miRNA-155, miRNA-210, and miRNA-20b is exaggerated in psoriasis and they may be involved in disease pathogenesis. A possible relationship between miRNA-210 and IL-17 may be suggested; however, further studies are still needed to verify this relation.

scholarly journals Evaluation of the relationship between IL-12, IL-13 and TNF-α gene polymorphisms with the susceptibility to brucellosis: a case control study

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Sima Kazemi ◽  
Asad Vaisi-Raygani ◽  
Fariba Keramat ◽  
Massoud Saidijam ◽  
Ali Reza Soltanian ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gene Polymorphism ◽  
Gene Polymorphisms ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Genotype Frequencies ◽  
Tnf Α ◽  
Polymerase Chain ◽  
The Relationship

Abstract Background The cytokine gene polymorphism is important for the genetic susceptibility of infectious diseases. The aim of the present study was to investigate the relationship between TNF-α, IL-12, and IL-13 gene polymorphisms and predisposition to brucellosis. Methods In this study, 107 patients with brucellosis and 107 healthy individuals were evaluated. The SNPs of TNF-α)- 238 G/A) and IL-12 (+ 1188 A/C) were done by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and IL-13 genotyping at positions − 1512 (A/C) and − 1112 (C/T) were analysis by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods. IL-12, IL-13 and TNF-α serum levels were measured by a sandwich enzyme-linked immunosorbent assay (ELISA). Results IL-13 (−1512A/C) was associated with Brucellosis risk in dominant model (OR (95% CI) = 2.17 (1.02–4.62)), P-value = 0.041. However, there was no difference in allele and genotype frequencies of TNF-α)- 238 G/A), IL-12 (+ 1188 A/C) and IL-13 [− 1512 (A/C) and − 1112 (C/T)] between patients and controls. Serum levels of IL-12 and TNF-α were significantly more frequent in the patients than in the control groups. Conclusions The IL-13 gene polymorphism can be used as a biomarker for detecting susceptibility to Brucella disease.

scholarly journals Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

scholarly journals Skin tissue expression and serum level of thymic stromal lymphopoietin in patients with psoriasis vulgaris

2019 ◽  
Vol 11 (1) ◽  
Author(s):  
Oki Suwarsa ◽  
Hartati Purbo Dharmadji ◽  
Endang Sutedja ◽  
Lengga Herlina ◽  
Putri Reno Sori ◽  
...  
Keyword(s):  
Psoriasis Vulgaris ◽  
Allergic Diseases ◽  
Serum Levels ◽  
Tissue Expression ◽  
Thymic Stromal Lymphopoietin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lesional Skin ◽  
Healthy Control ◽  
Polymerase Chain ◽  
The Difference

Thymic stromal lymphopoietin (TSLP) is known to be associated with allergic diseases. It is also suggested that TSLP has a role in autoimmune diseases such as psoriasis; however, the associated pathways remain unknown. There is currently little information on TSLP in psoriasis vulgaris. We investigated TSLP expressions on lesional and non-lesional skin of psoriasis vulgaris patients using reverse transcription- polymerase chain reaction. TSLP level was also investigated in serum from psoriasis vulgaris patients compared to healthy control using enzyme-linked immunosorbent assay. TSLP expression was higher in lesional skin (1.90) compared to non-lesional skin (1.76); however, the difference was not statistically significant (P>0.05). TSLP serum levels were significantly higher in psoriasis patients (287.40 pg/dL) as compared to controls (114.70 pg/dL) (P<0.05). This study concluded that TSLP levels in the serum of psoriasis vulgaris patients are higher than controls. TSLP was also found in keratinocyte of psoriasis patients, the expression was higher in the lesional compared to non-lesional skin; however, this difference is statistically insignificant. These findings suggest that TSLP may play a role in the pathogenesis of psoriasis vulgaris, but its exact role remains unclear.

Detection of small round structured viruses in clinical and environmental samples by polymerase chain reaction

1995 ◽  
Vol 31 (5-6) ◽  
pp. 375-382 ◽  
Author(s):  
M. Wolfaardt ◽  
C. L. Moe ◽  
W. O. K. Grabow
Keyword(s):  
Polymerase Chain Reaction ◽  
Tap Water ◽  
Practical Method ◽  
Glass Wool ◽  
Polluted Water ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Elution Method

Norwalk (NV) and other small round structured viruses (SRSVs) have been identified as common causes of gastroenteritis. Outbreaks of Norwalk gastroenteritis have been associated with contaminated drinking water and food such as oysters and salads. The cloning and sequencing of the NV genome has made it possible to detect NV and related viruses by the reverse transcription-polymerase chain reaction (RT-PCR). We applied RT-PCR to detect SRSVs in faecal specimens from two gastroenteritis outbreaks in South Africa, designated “Christmas” and “Grootbrak” and were able to detect SRSVs in all of the three specimens from the Christmas outbreak and in two of 16 specimens from the Grootbrak outbreak. The RT-PCR procedure used appeared to be more sensitive for the detection of SRSVs in patient stool specimens than immune electron microscopy and NV antigen detection by enzyme linked immunosorbent assay. The RT-PCR procedure proved suitable for the detection of SRSVs in seeded samples of sewage, sewage sludge, river water, and tap water. However, sensitivity was lower for seeded samples of sewage and sludge than for tap water, which indicates interference by high levels of organic matter. The RT-PCR procedure was also used to show that small numbers of SRSVs can successfully be recovered from large volumes of water by means of a glass wool adsorption-elution method. Since no practical method is available for quantitation of the small numbers of SRSVs concerned, it was not possible to evaluate the efficiency of recovery. Although no SRSVs have been detected by direct testing of sewage and sludge samples, the results obtained in this study show that RT-PCR detection of SRSVs in sewage and polluted water environments is feasible, and that small numbers of the viruses can, like many other enteric viruses, successfully be recovered by means of a glass wool adsorption-elution method.

scholarly journals Development of PCR-ELISA for Detection and Differentiation of Didymella bryoniae from Related Phoma species

Plant Disease ◽  
2002 ◽  
Vol 86 (7) ◽  
pp. 710-716 ◽  
Author(s):  
B. M. Somai ◽  
A. P. Keinath ◽  
R. A. Dean
Keyword(s):  
Gel Electrophoresis ◽  
Sulfonic Acid ◽  
Pcr Primers ◽  
Enzyme Linked Immunosorbent Assay ◽  
Didymella Bryoniae ◽  
Chain Reaction ◽  
Pcr Products ◽  
Elisa Technique ◽  
Polymerase Chain ◽  
Biotin Label

The causal agent of gummy stem blight, Didymella bryoniae, often is isolated from infected cucurbits together with other Phoma spp. Polymerase chain reaction (PCR) primers specific to D. bryoniae and Phoma were used to develop and evaluate a microtiter-based PCR-enzyme-linked immunosorbent assay (ELISA) technique. Primers were modified by addition of a fluorescein and a biotin label to the 5′ ends of the forward and reverse primers, respectively. After amplification, PCR products were detected in an ELISA using horseradish peroxidase-conjugated antifluorescein antibody and three substrates that yielded three colored products, one for each fungal group. The most sensitive substrate (highest signal:noise ratio) was 2,2′ -azino-bis[3-ethylbenz-thiazoline-6-sulfonic acid]. PCR-ELISA successfully detected 45 of 46 D. bryoniae and all 13 Phoma isolates that were used. Results were comparable to those obtained with gel electrophoresis. Only one D. bryoniae isolate could not be detected with PCR-ELISA; this isolate also produced a fragment larger than other D. bryoniae isolates on agarose gels. PCR-ELISA was used successfully on crude extracts of “blind” fungal samples and identified seven of seven isolates as D. bryoniae or Phoma. Although less sensitive than gel electrophoresis, PCR-ELISA was a highly specific, yet simple, rapid and convenient assay for detection of D. bryoniae and Phoma sp.

scholarly journals RT-PCR Method for Detecting Cowpea Mottle Carmovirus in Vigna Germ Plasm

Plant Disease ◽  
1999 ◽  
Vol 83 (7) ◽  
pp. 639-643 ◽  
Author(s):  
A. G. Gillaspie ◽  
S. E. Mitchell ◽  
G. W. Stuart ◽  
R. F. Bozarth
Keyword(s):  
Germ Plasm ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mild Mosaic ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Viral Pathogens ◽  
Highly Sensitive ◽  
Pcr Method ◽  
The Common ◽  
Polymerase Chain

A highly sensitive reverse transcription-polymerase chain reaction (RT-PCR) method was developed to detect cowpea mottle carmovirus (CPMoV) in newly acquired germ plasm of Vigna spp. It detected virus in tissues diluted up to 10-9. The preferred primers were designed from the RNA replicase cDNA sequence of CPMoV. These primers were able to detect CPMoV in plants infected with 10 different isolates of the virus. There were no cross-reactions with either bean mild mosaic or melon necrotic spot carmoviruses or any of the common cowpea viral pathogens tested. The RT-PCR method was up to 105 times more sensitive than direct antigen coating enzyme-linked immunosorbent assay (DAC-ELISA) in detecting CPMoV. The RT-PCR method gave no false positive reaction as is sometimes seen with ELISA.

scholarly journals Circulation of bluetongue virus in goats in the Karamoja region of Uganda

2013 ◽  
Vol 84 (1) ◽  
Author(s):  
Elijah N. Mulabbi ◽  
Chrisostom Ayebazibwe ◽  
Samuel Majalija ◽  
Carrie A. Batten ◽  
Christopher A.L. Oura
Keyword(s):  
Real Time ◽  
Whole Blood ◽  
Bluetongue Virus ◽  
Rna Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
High Seroprevalence ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Karamoja Region

The presence of bluetongue virus (BTV) in indigenous goats from the Karamoja region of northern Uganda was investigated. A total of 300 goats were sampled (serum and whole blood) from five districts within the Karamoja region. The samples were analysed for the presence of bluetongue (BT) antibodies using a commercial Enzyme-linked immunosorbent assay (ELISA) and for the presence of BTV viral RNA by real-time Reverse transcription polymerase chain reaction (RT-PCR), because BTV is an RNA virus. Of the 300 goats tested, 269 (90%) were positive for BTV antibodies, indicating high levels of BTV circulation within the region. Out of the 150 whole blood samples tested for the presence of the virus by real-time RT-PCR, 84 (56%) were positive for BTV RNA. This study, which is the first of its kind in Uganda, showed a high seroprevalence of BT antibodies and active circulation of BTV in a high proportion of goats in the Karamoja region.

scholarly journals Association between VEGF -2578 C> A Polymorphism with VEGF Serum Levels in Gastritis with Helicobacter Pylori

2021 ◽  
Vol 8 (8) ◽  
pp. 171-175
Author(s):  
Tara Rizvira Monica ◽  
Gontar Alamsyah Siregar ◽  
Taufik Sungkar
Keyword(s):  
Serum Levels ◽  
Mucosal Damage ◽  
Cross Sectional Study ◽  
Endoscopic Examination ◽  
Chain Reaction ◽  
Cross Sectional ◽  
Excess Production ◽  
Polymerase Chain ◽  
H Pylori ◽  
The Relationship

Mucosal damage in people with gastritis causes the production of VEGF. VEGF is a neoangiogenesis function to repair damaged tissue. Excess production can cause cancer risk. VEGF genotype polymorphisms are thought to affect the production of serum VEGF levels. The aim of this study was to determine the relationship between VEGF - 2578 C> A polymorphism and serum VEGF levels in H. pylori gastritis. Method: cross-sectional study was conducted at H. Adam Malik General Hospital and Network Hospital with 100 samples. Endoscopic examination was performed to assess the gastric mucosa and a tissue biopsy was performed. The urea breath test (UBT) test and the Campylobacter like organism (CLO) test to determine H. pylori infection. VEGF - 2578 C> A was checked by Polymerase Chain Reaction (PCR). The data will be analyzed by univariate and bivariate. Result: One hundred people with gastritis, of which 59 people were infected with H. pylori. In this study, H. pylori infection did not have a significant relationship with VEGF levels. VEGF - 2578 C> A polymorphisms also had no relationship to serum VEGF levels. Conclusion: There is no correlation between VEGF - 2578 C> A polymorphism with VEGF serum levels (p> 0.05). Keywords: VEGF polymorphisms, VEGF - 2578 CA, H. pylori, Gastritis.

Interlaboratory Evaluation of a New Reverse Transcriptase Polymerase Chain Reaction–Based Enzyme-Linked Immunosorbent Assay for the Detection of Circulating Melanoma Cells: A Multicenter Study of the Dermatologic Cooperative Oncology Group

2001 ◽  
Vol 19 (6) ◽  
pp. 1723-1727 ◽  
Author(s):  
U. Reinhold ◽  
C. Berkin ◽  
A.-K. Bosserhoff ◽  
A. Deutschmann ◽  
C. Garbe ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tumor Cells ◽  
High Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Blood Samples ◽  
Melanoma Patients ◽  
Polymerase Chain ◽  
Interlaboratory Reproducibility

PURPOSE: Reverse transcription-polymerase chain reaction (RT-PCR)–based detection of tyrosinase mRNA is the most frequently used laboratory method for the detection of circulating tumor cells in melanoma patients. However, previously published results showed considerable variability in the PCR positivity rates. MATERIALS AND METHODS: We designed a collaborative study to assess the sensitivity, specificity, and clinical relevance of a new standardized RT-PCR–based enzyme-linked immunosorbent assay (ELISA) for the detection of circulating melanoma cells. Blood samples of healthy donors mixed with cells of a melanoma cell line were prepared in a blinded fashion, and aliquots were sent to seven participating laboratories experienced in RT-PCR. RESULTS: The results demonstrate a high sensitivity (1 melanoma cell/mL blood) and specificity (no false-negatives and 7.4% [2 of 28] false-positives) of the assay and a satisfactory rate of interlaboratory reproducibility. The analysis of aliquots of blinded samples derived from 60 melanoma patients identified tyrosinase mRNA in 17 of 60 (28.3%): three (20%) of 15 stage I patients, two (13.3%) of 15 stage II patients, five (35.7%) of 14 stage III patients, and seven (43.8%) of 16 stage IV patients. The interlaboratory reproducibility of positive samples, however, was extremely low and indicates the presence of low amounts of target mRNA. CONCLUSION: Reverse transcriptase-PCR ELISA has a high sensitivity and specificity for the detection of tyrosinase mRNA in peripheral blood cells. The low interlaboratory reproducibility for the detection of tumor cells in blood samples of melanoma patients, however, raises the question of relevance of this assay for clinical use.

scholarly journals Evaluation of peanut genotypes for resistance to Tomato spotted wilt virus by mechanical and thrips inoculation

2006 ◽  
Vol 41 (6) ◽  
pp. 937-942 ◽  
Author(s):  
Luciana Cordeiro do Nascimento ◽  
Viboon Pensuk ◽  
Nivânia Pereira da Costa ◽  
Francisco Miguel de Assis Filho ◽  
Gilvan Pio-Ribeiro ◽  
...  
Keyword(s):  
Tomato Spotted Wilt Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mechanical Inoculation ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Breeding Lines ◽  
Tomato Spotted Wilt ◽  
Ic 10 ◽  
Polymerase Chain ◽  
Wilt Virus

The objective of this work was to evaluate the reactions of three peanut breeding lines (IC-10, IC-34, and ICGV 86388) to Tomato spotted wilt virus (TSWV) by mechanical and thrips inoculation, under greenhouse conditions, and compare them to the reactions of cultivars SunOleic, Georgia Green, and the breeding line C11-2-39. TSWV infection by mechanical inoculation was visually assessed using an index ranging from 0 (no symptoms) to 4 (apical death). Enzyme-linked immunosorbent assay was used to confirm TSWV infection from both mechanical and thrips inoculations. IC-10, IC-34, ICGV 86388, and C11-2-39 were more resistant than the cultivars SunOleic and Georgia Green based on mechanical inoculation. Upon thrips inoculation only IC-34 and ICGV-86388 were infected by TSWV, as demonstrated by reverse transcription polymerase chain reaction (RT-PCR), although no symptoms of infection were observed. The peanut breeding lines IC-10, IC-34, and ICGV 86388 show higher level of resistance to TSWV than cultivar Georgia Green considered a standard for TSWV resistance.

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