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

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.

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Detection of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in clinical samples using Real-Time Reverse Transcription Polymerase Chain Reaction (qRT-PCR)

IOP Conference Series Earth and Environmental Science ◽

10.1088/1755-1315/762/1/012026 ◽

2021 ◽

Vol 762 (1) ◽

pp. 012026

Author(s):

A Atikana ◽

L Sukmarini ◽

Hariyatun ◽

A M Ridwanulah ◽

H A Nugroho ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Severe Acute Respiratory Syndrome ◽

Real Time ◽

Reverse Transcription ◽

Clinical Samples ◽

Chain Reaction ◽

Qrt Pcr ◽

Polymerase Chain

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Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

Virology Journal ◽

10.1186/s12985-020-01467-y ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Yang Zhang ◽

Chunyang Dai ◽

Huiyan Wang ◽

Yong Gao ◽

Tuantuan Li ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Clinical Samples ◽

Chain Reaction ◽

Pcr Assay ◽

Qrt Pcr ◽

Highly Sensitive ◽

One Step ◽

Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

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Helicobacter pylori clarithromycin resistance detected by Etest and TaqMan real-time polymerase chain reaction: a comparative study

Apmis ◽

10.1111/j.1600-0463.2012.02896.x ◽

2012 ◽

Vol 120 (9) ◽

pp. 712-717 ◽

Cited By ~ 21

Author(s):

Rosa Monno ◽

Floriana Giorgio ◽

Panella Carmine ◽

Leonardo Soleo ◽

Vittoria Cinquepalmi ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Helicobacter Pylori ◽

Comparative Study ◽

Real Time ◽

Chain Reaction ◽

Clarithromycin Resistance ◽

Polymerase Chain

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An improved quantitative real-time polymerase chain reaction technology for Helicobacter pylori detection in stomach tissue and its application value in clinical precision testing

BMC Biotechnology ◽

10.1186/s12896-020-00624-z ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Ling Deng ◽

Xiao-Yi He ◽

Bin Tang ◽

Yang Xiang ◽

Juan-Juan Yue

Keyword(s):

Polymerase Chain Reaction ◽

Helicobacter Pylori ◽

Real Time ◽

Chain Reaction ◽

Stomach Tissue ◽

Polymerase Chain Reaction Technology ◽

Polymerase Chain ◽

Precision Testing

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Quantitation of 35S Promoter in Maize DNA Extracts from Genetically Modified Organisms Using Real-Time Polymerase Chain Reaction, Part 2: Interlaboratory Study

Journal of AOAC International ◽

10.1093/jaoac/88.2.558 ◽

2005 ◽

Vol 88 (2) ◽

pp. 558-573 ◽

Cited By ~ 18

Author(s):

Max Feinberg ◽

Sophie Fernandez ◽

Sylvanie Cassard ◽

Chrystèle Charles-Delobel ◽

Yves Bertheau ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Genetically Modified Organisms ◽

Genetically Modified ◽

Interlaboratory Study ◽

Performance Criteria ◽

Tolerance Interval ◽

Chain Reaction ◽

Qrt Pcr ◽

Polymerase Chain

Abstract The European Committee for Standardization (CEN) and the European Network of GMO Working Laboratories have proposed development of a modular strategy for stepwise validation of complex analytical techniques. When applied to the quantitation of genetically modified organisms (GMOs) in food products, the instrumental quantitation step of the technique is separately validated from the DNA extraction step to better control the sources of uncertainty and facilitate the validation of GMO-specific polymerase chain reaction (PCR) tests. This paper presents the results of an interlaboratory study on the quantitation step of the method standardized by CEN for the detection of a regulatory element commonly inserted in GMO maize-based foods. This is focused on the quantitation of P35S promoter through using the quantitative real-time PCR (QRT-PCR). Fifteen French laboratories participated in the interlaboratory study of the P35S quantitation operating procedure on DNA extract samples using either the thermal cycler ABI Prism® 7700 (Applied Biosystems, Foster City, CA) or Light Cycler® (Roche Diagnostics, Indianapolis, IN). Attention was focused on DNA extract samples used to calibrate the method and unknown extract samples. Data were processed according to the recommendations of ISO 5725 standard. Performance criteria, obtained using the robust algorithm, were compared to the classic data processing after rejection of outliers by the Cochran and Grubbs tests. Two laboratories were detected as outliers by the Grubbs test. The robust precision criteria gave values between the classical values estimated before and after rejection of the outliers. Using the robust method, the relative expanded uncertainty by the quantitation method is about 20% for a 1% Bt176 content, whereas it can reach 40% for a 0.1% Bt176. The performances of the quantitation assay are relevant to the application of the European regulation, which has an accepted tolerance interval of about ±50%. These data were fitted to a power model (r2 = 0.96). Thanks to this model, it is possible to propose an estimation of uncertainty of the QRT-PCR quantitation step and an uncertainty budget depending on the analytical conditions.

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Prevalence of the Helicobacter pylori babA2 Gene in Children Mainly Depends on the PCR Primer Set Used

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2020/4080248 ◽

2020 ◽

Vol 2020 ◽

pp. 1-9

Author(s):

Anja Šterbenc ◽

Maja M. Lunar ◽

Matjaž Homan ◽

Boštjan Luzar ◽

Nina Zidar ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Helicobacter Pylori ◽

Diagnostic Tool ◽

Clinical Relevance ◽

Detection Methods ◽

Chain Reaction ◽

Polymerase Chain ◽

Primer Sets ◽

H Pylori ◽

Pcr Primer

Various polymerase chain reaction- (PCR-) based methods with varying positivity rates were designed to detect the Helicobacter pylori babA2 gene. To compare different primer sets, babA2 prevalence was determined in 279 H. pylori-positive pediatric samples using the 832 bp, 139 bp, and 271 bp PCR primer sets, resulting in 34.0%, 51.3%, and 79.6% prevalence of the babA2 gene, respectively. The babA2 status determined using the 832 bp and 139 bp PCR primer sets significantly correlated with bacterial density and activity of inflammation, whereas no such correlations were found using the 271 bp PCR primer set. The 139 and 832 bp PCR primer sets concordantly detected the babA2 gene in 93 cases; however, in comparison to the 832 bp PCR primer set, the 139 bp PCR primer set detected additional 50 babA2 cases, whereas only two 832 bp positive cases were missed. The 271 bp PCR primer set missed 32 babA2 cases that were 832 bp and/or 139 bp PCR positive, but tested solely positive in 109 cases. Interestingly, cloning of a subset of 271 bp PCR positive samples revealed amplification of the babA/B gene chimera. Hence, in our opinion, the 271 bp PCR protocol is not a reliable diagnostic tool for detecting the babA2 gene in children. Our results reaffirm previous observations that the use of certain babA2 PCR primer sets can significantly impact estimation of the prevalence and clinical relevance of the H. pylori babA2 gene in children, suggesting babA2 detection methods should be carefully selected.

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