Mycoplasma pneumoniaeandChlamydophila pneumoniae: a comparative study in patients with nasal polyposis and healthy controls

2015 ◽  
Vol 129 (9) ◽  
pp. 865-869 ◽  
Author(s):  
D G Ioannidis ◽  
V A Lachanas ◽  
Z Florou ◽  
J G Bizakis ◽  
E Petinaki ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Inferior Turbinate ◽  
Sinus Surgery ◽  
Control Group ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain

AbstractIntroduction:The role played byMycoplasma pneumoniaeandChlamydophila pneumoniaein the pathogenesis of chronic rhinosinusitis with nasal polyps has been the object of ongoing debate. We used real-time polymerase chain reaction to investigate the prevalence of both microorganisms in the nasal tissue samples of patients and controls.Methods:We extracted DNA from nasal polyp samples obtained during functional endoscopic sinus surgery and the inferior turbinate samples of controls undergoing septoplasty. We used the highly sensitive real-time polymerase chain reaction to detect the presence ofM pneumoniaeandC pneumoniaeDNA.Results:Patients with chronic rhinosinusitis with nasal polyps consisted of 62 individuals (39 men; mean age 51 years); the control group consisted of 24 individuals (13 men; mean age 45 years). All samples from both groups were negative forM pneumoniaeandC pneumoniaeDNA.Conclusion:We have demonstrated that the likelihood ofM pneumoniaeandC pneumoniaeacting as an ongoing inflammatory stimulus in chronic rhinosinusitis with nasal polyps is slim.

scholarly journals Pilot study of DNA methylation in the pathogenesis of chronic rhinosinusitis with nasal polyps

2015 ◽  
Vol 53 (4) ◽  
pp. 345-352
Author(s):  
Y.B. Zheng ◽  
Y. Zhao ◽  
L.Y. Yue ◽  
P. Lin ◽  
Y.F. Liu ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Polymerase Chain Reaction ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Cpg Islands ◽  
Inferior Turbinate ◽  
Control Group ◽  
Chain Reaction ◽  
Bisulphite Sequencing ◽  
Polymerase Chain

Background: DNA methylation has been implicated in the pathogenesis of allergy and atopy. This study aimed to identify whether DNA methylation also plays an important role in the pathogenesis of nasal polyps (NP). Methodology: NP tissues were obtained from 32 patients with chronic rhinosinusitis with bilateral NP. Biopsies of inferior turbinate mucosa (ITM) were taken from 18 patients who underwent rhinoseptoplasty (control group). The methylated genes, which were detected by DNA methylation microarray, were validated by methylation-specific polymerase chain reaction, bisulphite sequencing, real-time polymerase chain reaction and immunohistochemistry. Results: DNA methylation microarray identified 8,008 CpG islands in 2,848 genes. One hundred and ninety-eight genes were found to have a methylated signal in the promoter region in NP samples compared with ITM samples. The four top genes that changed, COL18A1, EP300, GNAS and SMURF1, were selected for further study. The methylation frequency of COL18A1 was significantly higher in NP samples than in ITM samples. Conclusions: DNA methylation might play an important role in the pathogenesis of NP. Promoter methylation of COL18A1 was found to be significantly increased in NP tissues, further studies are necessary to confirm the significance of these epigenetic factors in the mechanisms underlying the development or persistence of NP.

scholarly journals Development of Quantitative Real-Time Polymerase Chain Reaction for Detection of and Discrimination between Erysipelothrix Rhusiopathiae and Other Erysipelothrix Species

2009 ◽  
Vol 21 (5) ◽  
pp. 701-706 ◽  
Author(s):  
Ho To ◽  
Tomohiro Koyama ◽  
Shinya Nagai ◽  
Kotaro Tuchiya ◽  
Tetsuo Nunoya
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Limit Of Detection ◽  
Enrichment Culture ◽  
Bacterial Species ◽  
Erysipelothrix Rhusiopathiae ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Practical Applications ◽  
Polymerase Chain

Quantitative real-time polymerase chain reaction (qPCR) assays were developed and validated in combination with enrichment culture for the detection and discrimination of Erysipelothrix rhusiopathiae and other Erysipelothrix species from tissue samples. The targets for SYBR green qPCR assays were the 16S ribosomal RNA gene for Erysipelothrix species and a gene involved in capsular formation for E. rhusiopathiae. The specificity of the assays was assessed with Erysipelothrix species and other related bacterial species. The limit of detection was found to be 5 colony-forming units per reaction. Amplification of DNA extracted from spleen and joint samples spiked with increasing quantities of Erysipelothrix cells was shown to be equally sensitive to DNA extracted from a pure bacterial culture. The assays were evaluated with 88 tissue samples from 3 experimentally infected pigs and 50 mice and with 36 tissue samples from 3 naturally infected pigs and 11 noninfected pigs. Results were compared with those of direct qPCR and conventional culture. The qPCR after enrichment increased the diagnostic sensitivity over that of culture and qPCR, thereby significantly reducing the total time taken for the detection of E. rhusiopathiae and other Erysipelothrix species. Therefore, this technique could be used for practical applications.

scholarly journals Aberrant UBR4 expressions in Hirschsprung disease patients

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Gunadi ◽  
Alvin Santoso Kalim ◽  
Estelita Liana ◽  
Aditya Rifqi Fauzi ◽  
Dian Nirmala Sirait ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Anorectal Malformation ◽  
E3 Ligase ◽  
Hirschsprung Disease ◽  
Control Group ◽  
Chain Reaction ◽  
Expression Change ◽  
Polymerase Chain ◽  
Hscr Patient

Abstract Background Recently, pathogenic alleles within ubiquitin N-recognin domain-containing E3 ligase 4 (UBR4) gene have been shown to be associated with Hirschsprung disease (HSCR). We determined the UBR4 expressions in Indonesian HSCR patients. Methods We analyzed the UBR4 expressions in the colons of HSCR patient and anorectal malformation (ARM) patient as control by real-time polymerase chain reaction (qPCR). Results Thirty-seven patients with non-syndromic HSCR and eighteen controls were involved in this study. qPCR revealed that the UBR4 expression was strongly decreased (0.77-fold) in the ganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.43 ± 0.36 vs. 2.05 ± 0.69; p = 0.009), whereas the UBR4 expression was also significantly reduced (0.79-fold) in the aganglionic group of patients with HSCR compared to the control group with ARM (ΔCT 2.39 ± 0.46 vs. 2.05 ± 0.69; p = 0.044). However, the UBR4 expression change was not associated with gender (p = 0.35 and 0.80), nor with degree of aganglionosis both in ganglionic and aganglionic colons (p = 0.72 and 0.73), respectively. Conclusion Our study demonstrates that expression of UBR4 is decreased in both aganglionic and ganglionic colon of HSCR patients.

Distinguishing Molecular Features of Allergic Fungal Rhinosinusitis

2018 ◽  
Vol 159 (1) ◽  
pp. 185-193 ◽  
Author(s):  
Matthew A. Tyler ◽  
Caroline J. Padro Dietz ◽  
Chris B. Russell ◽  
Martin J. Citardi ◽  
Shervin Assassi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Rhinosinusitis ◽  
Quantitative Polymerase Chain Reaction ◽  
Sinus Surgery ◽  
Type I ◽  
Chain Reaction ◽  
Molecular Features ◽  
Fungal Rhinosinusitis ◽  
Allergic Fungal Rhinosinusitis ◽  
Polymerase Chain

Objective Allergic fungal rhinosinusitis (AFRS) is a clinical subtype of chronic rhinosinusitis with nasal polyps (CRSwNP), characterized by eosinophilic mucin, evidence of fungal elements within the mucin, fungal-specific type I hypersensitivity, and characteristic computed tomography findings. It remains controversial whether AFRS represents a disease with a unique pathophysiology from chronic rhinosinusitis or is merely a severe form of CRSwNP. The goal of this study was to identify molecular features unique to AFRS. Study Design Cross-sectional case-control. Setting Single academic tertiary referral institution. Subjects and Methods Subjects included 86 patients undergoing endoscopic sinus surgery: CRSwNP (n = 34), AFRS (n = 37), and healthy controls (n = 15). Pathway and correlation analyses were performed with whole-genome microarray data for study patients undergoing surgery for recalcitrant chronic rhinosinusitis. Our findings were confirmed with quantitative polymerase chain reaction and immunohistochemical studies. Results AFRS was uniquely characterized by a pronounced association with adaptive T helper 2–associated immune gene expression. AFRS exhibited altered expression of proteins associated with secretory salivary peptides—namely, histatin, a peptide with known antifungal activity in the oral cavity. Furthermore, the expression of histatins correlated negatively with that of type 2 inflammatory mediators. We confirm the decreased expression of histatins in AFRS when compared with CRSwNP by quantitative polymerase chain reaction and localized its expression to a submucosal cell population. Conclusion There exist clear molecular profiles that distinguish AFRS from CRSwNP. This divergence translates into an altered ability to control fungal growth and may in part explain some of the phenotypical differences between CRSwNP and AFRS.

scholarly journals Investigation of the role of major respiratory viruses in the aetiology of nasal polyps using polymerase chain reaction technique

2014 ◽  
Vol 128 (4) ◽  
pp. 356-359
Author(s):  
F Aksoy ◽  
A Yenigun ◽  
R Dogan ◽  
F Yilmaz ◽  
O Ozturan ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Nasal Polyps ◽  
Nasal Polyposis ◽  
Respiratory Viruses ◽  
Polymerase Chain Reaction Technique ◽  
Control Group ◽  
Chain Reaction ◽  
Human Coronavirus ◽  
Polymerase Chain

AbstractObjective:We aimed to identify the role of major respiratory viruses in the aetiology of human nasal polyps using polymerase chain reaction technique.Methods:Thirty patients with nasal polyps and a group of 20 healthy patients (control group) were included in this study. Mucosa was obtained from the polyps of patients with nasal polyposis and from the middle turbinate of the control group patients by means of biopsy. The samples were stored at −80 °C until molecular analysis by polymerase chain reaction was carried out.Results:In the control group, the human coronavirus and human rhinovirus were diagnosed in one of the patients and the human respiratory syncytial virus in another. In the group with nasal polyposis, the influenza B virus was identified in one of the patients and the human coronavirus in another.Conclusion:The results did not demonstrate a statistically significant relationship between nasal polyposis and respiratory viruses.

scholarly journals FN1 mRNA expression of fibronectin 1 and distribution of fibronectin-associated leukocytes in humans with chronic diffuse liver diseases

2020 ◽  
Vol 11 (2) ◽  
pp. 200-206
Author(s):  
H. V. Dolhikh ◽  
H. S. Maslak ◽  
G. P. Chernenko ◽  
О. H. Minchenko ◽  
А. О. Dolhikh
Keyword(s):  
Polymerase Chain Reaction ◽  
Mrna Expression ◽  
Real Time ◽  
Liver Diseases ◽  
Support Vector ◽  
Control Group ◽  
Chain Reaction ◽  
Prognostic Accuracy ◽  
Polymerase Chain ◽  
Diffuse Liver Diseases

Chronic diffuse liver diseases are characterized by continuous progression of fibrosis, ultimately leading to cirrhosis with the following loss of the normal functioning of this organ due to excessive accumulation of the components of extracellular matrix. To find new, more available diagnostic markers of detecting disorders in the liver, we used methods of antifungal cytofluorometry and quantitative real-time polymerase chain reaction. Intensity of exposure of fibronectin and plasmatic membrane of lymphocytes in the group of patients with chronic diffuse diseases compared with the control group of practically healthy donors decreased both inside and on the surface of the cells respectively by 45.3% and 16.2%. Similar tendency towards decrease was observed during the assays of the level of the exposure of fibronectin on the surface and inside the blood granulocytes: by 25.0% and 36.5%, respectively. In the blood of the patients suffering from chronic diffuse diseases, compared with the control group, there was determined reliable increase in percentage of lymphocytes and granulocytes which contain topical fibronectin, by 32.3% and 2.78 times, correspondingly. The level of monocytes (as a percentage) with cell-associated fibronectin and fibronectin localized inside, by contrast, reliably decreased in 2.07 and 4.50 times, respectively. Analysis of the expression of FN1 in lymphocytes of blood of the studied groups using quantitative real-time polymerase chain reaction revealed decrease in the level of FN1 mRNA expression by 34.0% in the group of ill patients compared with the control group. We determined excellent diagnostic informativeness of the parameters of the level of exposure of fibronectin inside and on the surface of granulocytes and prognostic accuracy of the classifier from these parameters at the level of 100% using the method of support vector machine, SVM. High levels of diagnostic informativeness were recorded for the tests of all types of analyzed leukocytes with cell-associated fibronectin, and the classifiers based on the pair combinations of the tests with cell-associated fibronectin and fibronectin localized within the cells provide high diagnostic accuracy of the prognosis. Because the mentioned indicators are highly-sensitive tests, they can be proposed for early diagnostics and evaluation of the effectiveness of the conducted therapy of chronic diffuse liver diseases, which would allow reducing the use of paracentetic trepanobiopsy, a painful and risky procedure, which still remains the main type of diagnostic.

scholarly journals Investigation of SP1 and SP3 Expressions in Colorectal Cancer Carcinogenesis

2021 ◽  
Vol 21 (03) ◽  
Author(s):  
Turkan Gurer
Keyword(s):  
Colorectal Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Pathological Features ◽  
Chain Reaction ◽  
Tissue Samples ◽  
High Positive Correlation ◽  
Pathological Characteristics ◽  
Significant Difference ◽  
Polymerase Chain

ABSTRACT The study aimed to determine the expressions of SP1 and SP3 and the clinic pathological characteristics of patients, and the correlation between the expressions of SP1 and SP3 in colorectal cancer (CRC). In this study, tumour and adjacent non-tumour tissue samples were obtained from 41 individuals with CRC. SP1 and SP3 expressions were performed using Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR). According to the results of our study, there was no statistically significant difference in SP1 and SP3 expression levels between tumour tissues and non-tumour tissues (p>0.05), as well as no association with clinic pathological features of patients. In addition, a high positive correlation was found between the expressions of SP1 and SP3 genes in CRC (p=0.01). Consequently, it can be said that there is a correlation between SP1 and SP3 expressions, but SP1 and SP3 expressions are not related to CRC carcinogenesis.

scholarly journals Refluks Helicobacter pylori di mukosa hidung penderita rinosinusitis kronik disertai refluks laringofaring

2019 ◽  
Vol 48 (2) ◽  
pp. 148
Author(s):  
Sinta Sari Ratunanda ◽  
Billy Talakua ◽  
Teti Madiadipoera ◽  
Thaufiq Boesoirie ◽  
Ratna Anggraeni ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Helicobacter Pylori ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Nasal Mucosa ◽  
Nasal Discharge ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
H Pylori

Latar belakang: Rinosinusitis kronik masih menjadi problema di seluruh dunia. Faktor yang berasosiasi dengan Rinosinusitis Kronik (RSK) diduga multifaktorial, salah satunya adalah refluks laringofaring (RLF). Isi refluks cairan lambung antara lain adalah bakteri Helicobacter pylori (H. pylori) yang dengan patomekanisme refluks, diduga dapat mencapai mukosa laringofaring bahkan sampai mukosa sinonasal, dan menyebabkan RSK. Tujuan: Mendeteksi  H. pylori di mukosa hidung akibat refluks pada penderita RSK disertai RLF. Bila terdeteksi H. pylori, tata laksana harus lebih komprehensif, sehingga diharapkan RSK menjadi terkontrol. Metode: Penelitian deskriptif untuk mengetahui ada tidaknya H. pylori di mukosa sinonasal penderita RSK dengan RLF. Deteksi H. pylori menggunakan teknik quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) dari bahan penyikatan mukosa hidung. Hasil: Didapatkan 86 orang penderita RSK disertai RLF, terdiri dari 30 (35%) pasien laki-laki dan 56 (65,0%) pasien wanita, dengan rerata usia 43,25±6,30 tahun. Keluhan RSK terbanyak adalah hidung tersumbat dengan skor VAS > 7 sebesar 76,8%. Skor nasoendoskopi RSK terbesar pada skor 2 untuk edema mukosa sebesar 65,3% dan skor 2 untuk sekret hidung sebesar 58,2%. Rata-rata skor gejala refluks (SGR) adalah 26,43±4,03 dan rata-rata total skor temuan refluks (STR) adalah 11,28±1,21. Hasil pemeriksaan deteksi H. pylori dengan qRT-PCR, 100% tidak menemukan H. pylori dari penyikatan mukosa hidung. Kesimpulan: Refluks berupa H. pylori tidak ditemukan pada mukosa hidung  penderita RSK disertai RLF. Penelitian lebih lanjut diperlukan dengan menggunakan gabungan beberapa metode pemeriksaan  bersamaan untuk deteksi H. pylori akibat refluks di mukosa sinonasal  penderita RSK disertai RLF.  Background: Chronic rhinosinusitis is presently still a worldwide problem. Assosiating factors  to chronic rhinosinusitis (CRS) are multifactorial, one of them is laryngopharyngeal reflux (LPR). The gastric juice contains Helicobacter pylori (H. pylori), which by pathologic reflux could reach laryngopharyngeal and sinonasal area causing CRS. Purpose: To detect H. pylori in nasal mucosa caused by reflux, which suspected of causing CRS with LPR disease. Should H. pylori be found in nasal mucosa, the management of the disease must be comprehensive to enable  controlling CRS. Methods: A descriptive study to detect H. pylori in nasal mucosa CRS with LPR patients, using Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) through nasal brushing. Results: Eighty-six CRS with LPR patients as study objects consisted of 30 (35%) male, and 56 (65%) female, the age mean was 43.25±6.3 years old. Visual Analoque Scale (VAS) score for nasal obstruction more than 7 was the highest complaint (76.8%). Nasal endoscopic score of mucosal edema (65.3%) and nasal discharge (58,2%) had score 2. The average total score reflux symptom index (RSI) was 26.43±4.03 and the total score reflux finding score (RFS) was 11.28±1.21. H. pylori detection found negative 100% in CRS with LPR specimens. Conclusion: This study did not find reflux containing H. pylori in nasal mucosa of CRS with LPR patients.    Suggesting further study using simultaneously several methods to detect H. pylori in nasal mucosa  CRS with LPR patients.

scholarly journals Comparative evaluation of a competitive polymerase chain reaction (PCR) and a SYBR Green–based real-time PCR to quantify Porcine circovirus-2 DNA in swine tissue samples

2011 ◽  
Vol 23 (6) ◽  
pp. 1160-1167 ◽  
Author(s):  
Diogenes Dezen ◽  
Franciscus A.M. Rijsewijk ◽  
Thais F. Teixeira ◽  
Carine L. Holz ◽  
Ana P. Varela ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Manifestations ◽  
Sybr Green ◽  
Porcine Circovirus ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Porcine Circovirus 2 ◽  
Polymerase Chain

Porcine circovirus-2 (PCV-2) is considered the major etiological agent of post-weaning multisystemic wasting syndrome (PMWS) in pigs. The clinical manifestations of the disease are correlated with moderate to high amounts of PCV-2 DNA in biological samples of affected pigs. A threshold of 107 DNA copies/ml is suggested as the trigger factor for symptoms. A comparative study was conducted to determine which quantitative method would be more suitable to estimate the PCV-2 DNA load. Two polymerase chain reaction (PCR) assays were developed: a competitive PCR (cPCR) and a SYBR Green–based real-time PCR. The assays were compared for their capacity to detect PCV-2 in DNA samples extracted from liver, lung, spleen, mesenteric lymph nodes, and kidney of PMWS-affected ( n = 23) or non–PMWS-affected pigs ( n = 9). Both assays could successfully quantify PCV-2 DNA in all tissue samples and were able to detect significant differences between the numbers of PCV-2 DNA copies found in tissues of PMWS-affected and non–PMWS-affected pigs (≥102.5). The highest mean viral loads were detected by the SYBR Green real-time PCR, up to 107.0±1.5 copies/100 ng of total DNA sample, while the cPCR detected up to 104.8±1.5. A mean difference of 101.8 was found between the amounts of PCV-2 DNA detected, using the SYBR Green real-time PCR and the cPCR, suggesting that the viral load threshold for PMWS should be determined for each particular assay.

Molecular Study of Cytomegalovirus Infection among Children with End Stage Renal Diseases Undergoing Dialysis, Pilot Study

2017 ◽  
Vol 2 (4) ◽  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Immunoglobulin M ◽  
Renal Diseases ◽  
Control Group ◽  
Chain Reaction ◽  
Specific Test ◽  
Polymerase Chain ◽  
End Stage

Cytomegalovirus is considered as an opportunistic infection affecting immunocompromized patients. Children with end stage renal diseases requiring dialysis is among affected population by this virus. The aim of the present study was to detect and compare the seroprevalence of CMV and CMV antigen pp65 with real time polymerase chain reaction (PCR) among children with end stage renal diseases undergoing dialysis. The study is a prospective case - control study. The forty one patients included in the studied are registered in the hospital for regular dialysis waiting for renal transplantation. The study included forty one healthy controls with same age and gender distribution. Blood samples were obtained from studied children and subjected for determination of specific immunoglobulin M and G for CMV (IgM-CMV, IgG-CMV) by Elecys system and CMV-DNA determination by real time polymerase chain reaction (PCR) and for PP65 antigenaemia test by light diagnostic CMVpp65. CMV-IgM was significantly detected frequently (P=0.0001) in 12.2% of the patients and in 2.4% of the control children. Moreover, IgG-CMV was significantly more frequently detected in patients (P=0.0001) than in control (90.2%&31.7% respectively). CMV-DNA was significantly (P=0.0001) detected in 12 patients (29.3%) compared to the control (2.4%), while CMVpp65 was detected among 4 children (9.8%) compared to one child in the control group. The comparison between IgM-CMV and real time PCR revealed that 30.7% of positive samples by PCR had positive IgMCMV, while IgG-CMV was associated with 84.6% of positive PCR. CMVpp65 correctly identified all negative samples compared to PCR, while the majority of negative PCR was also negative for IgM-CMV (98.6%). Moreover, all negative children for CMVpp65 was also negative by PCR (100%) For the validity of different CMV markers, IgG-CMV was the most sensitive test (84.7%), CMVpp65 was the most specific test 100%. From this study we concluded that CMV is a common viral infection among children with end stage renal diseases requiring dialysis. The diagnostic performance of real time PCR is the gold standard technique in diagnosis of this infection. CMVpp65 antigenemia is a specific accurate test for laboratory diagnosis however, it lacks sensitivity. Specific IgG for CMV is good screening diagnostic test.

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