RESUMO
AIM: To develop the simple, rapid and sensitive dual-label time-resolved fluoroimmunoassay for pepsinogens in human serum. METHODS: Based on two-site sandwich protocol, monoclonal antibodies (McAbs) against pepsinogen I (PG I) and PG II were co-coated in 96 microtitration wells, and tracer McAbs against PG I and PG II were labeled with europium (Eu) and samarium (Sm) chelate, respectively. Diluted serum samples of Eu(3+)- and Sm(3+)-McAbs were added into microtitration wells simultaneously. After washing, fluorescence of bound Sm(3+) and Eu(3+) tracers was detected. RESULTS: The detection limit was 0.2 microg/L for PG I and 0.05 microg/L for PG II. The assay range was 5.0-320.0 microg/L for PG I and 1.0-55.0 microg/L for PG II. The average recovery rate was 102.7% for PG I and 98.8% for PG II. Sera from healthy controls and patients with gastric disease were analyzed. The PG detected by dual-label assay was in good agreement with that detected by single-label assay or by enzyme-linked immunosorbent assay. CONCLUSION: Dual-label assay can provide high-throughput serological screening for gastric diseases.
Assuntos
Fluorimunoensaio/métodos , Pepsinogênios/sangue , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/metabolismo , Ensaio de Imunoadsorção Enzimática/métodos , Európio/química , Fluorimunoensaio/normas , Gastroenteropatias/sangue , Gastroenteropatias/diagnóstico , Humanos , Limite de Detecção , Samário/química , Sensibilidade e EspecificidadeAssuntos
Adenocarcinoma Mucinoso/patologia , Neoplasias Pancreáticas/patologia , Adenocarcinoma Mucinoso/metabolismo , Adenocarcinoma Mucinoso/cirurgia , Idoso , Biomarcadores Tumorais/metabolismo , Antígeno Carcinoembrionário/metabolismo , Carcinoma Ductal Pancreático/metabolismo , Carcinoma Ductal Pancreático/patologia , Carcinoma Ductal Pancreático/cirurgia , Carcinoma Papilar/metabolismo , Carcinoma Papilar/patologia , Carcinoma Papilar/cirurgia , Cistadenocarcinoma Mucinoso/metabolismo , Cistadenocarcinoma Mucinoso/patologia , Cistadenocarcinoma Mucinoso/cirurgia , Diagnóstico Diferencial , Neoplasias Duodenais/metabolismo , Neoplasias Duodenais/patologia , Neoplasias Duodenais/cirurgia , Feminino , Seguimentos , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Mucina-2 , Mucinas/metabolismo , Invasividade Neoplásica , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/cirurgia , PancreaticoduodenectomiaRESUMO
OBJECTIVE: To explore the value of analysis of p53 gene mutation in fecal specimen in the diagnosis of colorectal carcinoma (CRC) in order to establish a non-invasive method for the screening of CRC. METHODS: The status of p53 gene mutation of the tumor tissues and corresponding fecal specimens was analyzed by polymerase chain reaction-single strand configuration polymorphism with EB staining in 40 CRC, 20 colorectal adenoma and 15 gastrocarcinoma. RESULTS: Amplification rates of fecal p53 gene were 90%, 85% and 93% in 40 CRC, 20 colorectal adenoma and 15 gastrocarcinoma, respectively. Total amplification rate was 89%. p53 gene mutation in tumor tissue was detected in 29 of 40 cases of CRC, 23 cases of which had p53 mutation in fecal specimens with the diagnostic sensitivity rate of 57.5%. Analysis of fecal p53 gene mutation had relatively higher diagnostic sensitivity rate than the detection of serum carcinoembryonic antigen and fecal occult blood (P < 0.05) for the diagnosis of CRC. 3 of 20 colorectal adenoma had p53 mutation both in tumor tissues and fecal specimens. 10 of 15 gastrocarcinoma had p53 mutation in tumor tissues but none in fecal specimens. CONCLUSIONS: Analysis of fecal p53 gene mutation has relatively higher diagnostic sensitivity rate for diagnosis of CRC and is expected to be a relatively sensitive, specific and effective method for early diagnosis of CRC, especially for CRC screening in large scale of the population.
