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胃蛋白酶(PP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法)

ELISA Kit for Pepsin (PP)

  • 胃蛋白酶(PP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 胃蛋白酶(PP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 產(chǎn)品包裝(模擬)
  • 胃蛋白酶(PP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) 實(shí)驗(yàn)結(jié)果圖
  • CEA632Ra.jpg 標(biāo)準(zhǔn)曲線圖
  • Certificate 通過ISO 9001、ISO 13485質(zhì)量體系認(rèn)證

特異性

本試劑盒用于檢測(cè)胃蛋白酶(PP),經(jīng)檢測(cè)與其它相似物質(zhì)無明顯交叉反應(yīng)。
由于受到技術(shù)及樣本來源的限制,不可能完成對(duì)所有相關(guān)或相似物質(zhì)交叉反應(yīng)檢測(cè),因此本試劑盒有可能與未經(jīng)檢測(cè)的其它物質(zhì)有交叉反應(yīng)。

回收率

分別于定值血清及血漿樣本中加入一定量的胃蛋白酶(PP)(加標(biāo)樣品),重復(fù)測(cè)定并計(jì)算其均值,回收率為測(cè)定值與理論值的比率。

樣本 回收率范圍(%) 平均回收率(%)
serum(n=5) 95-104 101
EDTA plasma(n=5) 86-95 90
heparin plasma(n=5) 86-95 90

精密度

精密度用樣品測(cè)定值的變異系數(shù)CV表示。CV(%) = SD/mean×100
批內(nèi)差:取同批次試劑盒對(duì)低、中、高值定值樣本進(jìn)行定量檢測(cè),每份樣本連續(xù)測(cè)定20 次,分別計(jì)算不同濃度樣本的平均值及SD值。
批間差:選取3個(gè)不同批次的試劑盒分別對(duì)低、中、高值定值樣本進(jìn)行定量測(cè)定,每個(gè)樣本使用同一試劑盒重復(fù)測(cè)定8次,分別計(jì)算不同濃度樣本的平均值及SD值。
批內(nèi)差: CV<10%
批間差: CV<12%

線性

在定值血清及血漿樣本內(nèi)加入適量的胃蛋白酶(PP),并倍比稀釋成1:2,1:4,1:8,1:16的待測(cè)樣本,線性范圍即為稀釋后樣本中胃蛋白酶(PP)含量的測(cè)定值與理論值的比率。

樣本 1:2 1:4 1:8 1:16
serum(n=5) 91-98% 79-89% 89-98% 85-99%
EDTA plasma(n=5) 89-101% 99-105% 92-105% 94-103%
heparin plasma(n=5) 80-90% 92-103% 99-105% 90-98%

穩(wěn)定性

經(jīng)測(cè)定,試劑盒在有效期內(nèi)按推薦溫度保存,其活性降低率小于5%。
為減小外部因素對(duì)試劑盒破壞前后檢測(cè)值的影響,實(shí)驗(yàn)室的環(huán)境條件需盡量保持一致,尤其是實(shí)驗(yàn)室內(nèi)溫度、濕度及溫育條件。其次由同一實(shí)驗(yàn)員來進(jìn)行操作可減少人為誤差。

實(shí)驗(yàn)流程

1. 實(shí)驗(yàn)前標(biāo)準(zhǔn)品、試劑及樣本的準(zhǔn)備;
2. 加樣(標(biāo)準(zhǔn)品及樣本)50µL,
    加入50µL檢測(cè)液A(臨用前配制);
    37°C溫育1小時(shí)。
3. 洗板3次;
4. 加檢測(cè)溶液B100µL,37°C孵育30分鐘;
5. 洗板5次;
6. 加TMB底物90µL,37°C孵育10-20分鐘;
7. 加終止液50µL,立即450nm讀數(shù)。

實(shí)驗(yàn)原理

本試劑盒應(yīng)用競(jìng)爭(zhēng)抑制酶聯(lián)免疫分析法測(cè)定標(biāo)本中待測(cè)物質(zhì)水平。將胃蛋白酶(PP)單克隆抗體包被微孔板,制成固相載體,往包被抗體的微孔中同時(shí)加入生物素標(biāo)記的抗原和待測(cè)抗原(標(biāo)準(zhǔn)品或樣本),待測(cè)抗原與生物素標(biāo)記抗原對(duì)特異性抗體進(jìn)行競(jìng)爭(zhēng)結(jié)合。溫育后經(jīng)洗滌去掉未結(jié)合物,然后加入HRP標(biāo)記的親和素,經(jīng)過溫育和徹底洗滌后加入底物TMB顯色。TMB在過氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成最終的黃色。待測(cè)標(biāo)本濃度越高,標(biāo)記抗原和抗體的結(jié)合就越受到抑制,顯色愈淺。顯色的深淺與酶量呈正相關(guān),而與樣品中待測(cè)物質(zhì)含量呈負(fù)相關(guān)。用酶標(biāo)儀在450nm波長(zhǎng)下測(cè)定吸光度(O.D.值),計(jì)算樣品濃度。

相關(guān)產(chǎn)品

編號(hào) 適用物種:Rattus norvegicus (Rat,大鼠) 應(yīng)用(僅供研究使用,不用于臨床診斷!)
CEA632Ra 胃蛋白酶(PP)檢測(cè)試劑盒(酶聯(lián)免疫吸附試驗(yàn)法) Enzyme-linked immunosorbent assay for Antigen Detection.
LMA632Ra 胃蛋白酶(PP)等多因子檢測(cè)試劑盒(流式熒光發(fā)光法) FLIA Kit for Antigen Detection.
KSA632Ra11 胃蛋白酶(PP)檢測(cè)試劑盒DIY材料(酶聯(lián)免疫吸附試驗(yàn)法) Main materials for "Do It (ELISA Kit) Yourself".

參考文獻(xiàn)

雜志 參考文獻(xiàn)
Radiology and Oncology LEVEL OF PEPSIN AND BILE ACIDS IN THE SALIVA OF PATIENTS WITH GLOTTIS T1 CARCINOMA [Szd:Source]
Radiology and Oncology Higher levels of total pepsin and bile acids in the saliva as a possible risk factor for early laryngeal cancer [Pubmed:Pmc4362607]
Clin Otolaryngol Pepsin and bile acids in saliva in patients with laryngopharyngeal reflux – a prospective comparative study [PubMed: 25516364]
journal of pediatric gastroenterology and nutrition Does Positioning Affect Tracheal Aspiration of Gastric Content in Ventilated Infants? [PubMed: 25313850]
Journal of Voice Detecting Laryngopharyngeal Reflux by Immunohistochemistry of Pepsin in the Biopsies of Vocal Fold Leukoplakia [pubmed:28756936]
Sensors (Basel) Label-Free Detection of Salivary Pepsin Using Gold Nanoparticle/Polypyrrole Nanocoral Modified Screen-Printed Electrode [Pubmed:29882917]
The Tohoku Journal of Experimental Medicine Association between Pepsin in Bronchoalveolar Lavage Fluid and Prognosis of Chronic Fibrosing Interstitial Lung Disease []
Respiratory?Research Pulmonary aspiration in preschool children with cystic fibrosis [Pubmed: 30558606]
Tanaffos Association of Bile Acid and Pepsin Micro-aspiration with Chronic Obstructive Pulmonary Disease Exacerbation [Pubmed: 31423141]
Sensors Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux [Pubmed: 31935973]
BIOSCIENCE REPORTS Presence of pepsin in laryngeal tissue and saliva in benign and malignant neoplasms [Pubmed: 33103719]
OTOLARYNGOLOGY-HEAD AND NECK SURGERY Relationship Between Pepsin Expression and Dysplasia Grade in Patients With Vocal Cord Leukoplakia [Pubmed: 32692278]
American Journal of Otolaryngology Association of pepsin and DNA damage in laryngopharyngeal reflux-related vocal fold polyps [Pubmed: 32889371]
The relationship between laryngopharyngeal reflux based on pepsin value and clinical characteristics of laryngeal cancer patients []
Med Sci Monit The Role of Salivary Pepsin in the Diagnosis of Gastroesophageal Reflux Disease (GERD) Evaluated Using High-Resolution Manometry and 24-Hour??- [33220027]
Annals of Esophagus Pepsin properties, structure, and its accurate measurement: a narrative review []
Laryngoscope The Impact of Laryngopharyngeal Reflux on Occurrence and Clinical Course of Recurrent Respiratory Papillomatosis [34338331]
Pulmonology Bronchoalveolar lavage (BAL) amylase and pepsin levels as potential biomarkers of aspiration pneumonia [Pubmed:35715334]
Neuromodulation: Technology at the Neural Interface Transcutaneous Electrical Acustimulation Improved the Quality of Life in Patients With Diarrhea-Irritable Bowel Syndrome [Pubmed:35088760]
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