Dataset Information

Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux.

ABSTRACT: Salivary pepsin is a promising marker for the non-invasive diagnosis of laryngopharyngeal reflux (LPR). For reliable results regarding pepsin in saliva, it is critical to standardize the collection, storage, and pre-processing methods. In this study, we optimized the saliva collection protocols, including storage conditions, i.e., solution, temperature, and time, and the pre-processing filter for pepsin. Moreover, we prepared a simple immunochromatographic strip for the rapid detection of pepsin and evaluated its sensing performance. As a result, we selected a polypropylene (PP) filter as the pre-processing filter for salivary pepsin in low resource settings, such as those where point of care testing (POCT) is conducted. This filter showed a similar efficiency to the centrifuge (standard method). Finally, we detected the pepsin using gold nanoparticles conjugated with monoclonal pepsin antibody. Under optimized conditions, the lower limit of detection for pepsin test strips was determined as 0.01 ?g/mL. Furthermore, we successfully detected the salivary pepsin in real saliva samples of LPR patients, which were pre-processed by the PP filter. Therefore, we expect that our saliva collection protocol and pepsin immunochromatographic strip can be utilized as useful tools for a non-invasive diagnosis/screening of LPR in POCT.

SUBMITTER: Lee YJ

PROVIDER: S-EPMC6982828 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux.

Lee Young Ju YJ Kwon Jiyoon J Shin Sanggyeong S Eun Young-Gyu YG Shin Jae Ho JH Lee Gi-Ja GJ

Sensors (Basel, Switzerland) 20200106 1

Salivary pepsin is a promising marker for the non-invasive diagnosis of laryngopharyngeal reflux (LPR). For reliable results regarding pepsin in saliva, it is critical to standardize the collection, storage, and pre-processing methods. In this study, we optimized the saliva collection protocols, including storage conditions, i.e., solution, temperature, and time, and the pre-processing filter for pepsin. Moreover, we prepared a simple immunochromatographic strip for the rapid detection of pepsin ...[more]

PMID: 31935973

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Project description:OBJECTIVE:Recently, a 24-h impedance was used to detect laryngopharyngeal reflux (LPR). However, not every case of LPR is pathological. Thus, pathological pharyngeal impedance values need to be clearly established to diagnose pathological LPR. The aim of our study was to establish pathological 24-h pharyngoesophageal impedance/pH values for the diagnosis of LPR. METHODS:The study was conducted in a tertiary care setting. A total of 30 patients who were referred to microlaryngoscopy for a laryngeal pathology that might be caused by LPR were included in this prospective study. All patients were off proton-pump inhibitor therapy. The 24-h pharyngoesophageal impedance-pH monitoring was performed 1 day before surgery. A biopsy of laryngeal tissue was obtained during microlaryngoscopy and was analyzed by immunohistochemistry to detect pepsin. The patients were divided into two groups: pepsin negative and pepsin positive (which indicated pathological LPR). The results of 24-h multichannel intraluminal impedance-dual-channel pH monitoring were compared between the groups. The number of LPR episodes in the pepsin-positive group was analyzed to establish a cutoff value for pathological LPR. RESULTS:There were 18 participants in the pepsin-negative group and 12 in the pepsin-positive group. The median total pharyngeal refluxes detected were two (0-5) in the pepsin-negative group and 14 (6-39) in the pepsin-positive group (P<0.001), although the groups were otherwise homogeneous. There was a statistically significant difference in the number of all types of refluxes between groups. Six or more pharyngeal refluxes were the cutoff for the presence of pepsin in the laryngeal mucosa and, thereby, for the diagnosis of relevant/pathological LPR. CONCLUSION:Six or more pharyngeal reflux episodes registered during the 24-h impedance/pH monitoring seem to be the cutoff for diagnosing pathological LPR. Therefore, it is possible to suggest establishing this value as the pathological impedance value indicating pathological LPR. These results must be interpreted with caution due to the small sample size.

Project description:OBJECTIVES:This study was conducted to investigate the current practices of Asian otolaryngologists for laryngopharyngeal reflux (LPR). METHODS:An online survey about LPR was sent to 2,000 members of Asian otolaryngological societies, and a subgroup analysis was performed between Western and Eastern Asian otolaryngologists. The survey was conducted by the Laryngopharyngeal Reflux Study Group of Young Otolaryngologists of the International Federation of Oto-rhino-laryngological Societies. RESULTS:Among approximately 1,600 Asian otolaryngologists, 146 completed the survey (62 from Western Asian countries, 84 from Eastern Asian countries). A substantial majority (73.3%) of the otolaryngologists considered LPR and gastroesophageal reflux disease to be different diseases. The symptoms thought to be closely related to LPR were coughing after lying down, throat clearing, and globus sensation. The findings thought to be closely related to LPR were posterior commissure granulations and hypertrophy, arytenoids, and laryngeal erythema. The respondents indicated that they mostly diagnosed LPR (70%) after an empirical therapeutic trial of proton pump inhibitors (PPIs). Although multichannel intraluminal impedance-pH (MII-pH) monitoring is a useful tool for diagnosing nonacid or mixed LPR, 78% of Asian otolaryngologists never or very rarely used MII-pH. Eastern Asian otolaryngologists more frequently used once-daily PPIs (64.3% vs. 45.2%, P=0.021), whereas Western Asian otolaryngologists preferred to use twice-daily PPIs (58.1% vs. 39.3%, P=0.025). The poor dietary habits of patients were considered to be the main reason for therapeutic failure by Asian otolaryngologists (53.8%). Only 48.6% of Asian otolaryngologists considered themselves to be adequately knowledgeable and skilled regarding LPR. CONCLUSION:Significant differences exist between Western and Eastern Asian otolaryngologists in the diagnosis and treatment of LPR. Future consensus statements are needed to establish diagnostic criteria and therapeutic regimens.

Dataset Information

Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux.

Publications

Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux.

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