Résumé
A molecular diagnostic assay which could be stored at room temperature was developed to rapidly detect Mycobacterium tuberculosis (MTB) based on loop-mediated isothermal amplification (LAMP) technology and dry-reagent process. LAMP uses 4 or 6 primers and Bst DNA polymerase to amplify DNA at a constant temperature. The results showed that the LAMP assay could detect the amplification of IS6110 target gene within 20 min using real-time fluorescence signal detection. The sensitive of LAMP assay was similar to the PCR technology while the precision of PCR was better than LAMP (coefficient of variation, LAMP 18.9%, PCR 3.4%), meaning LAMP was more suitable for qualitative detection. The LAMP assay did not amplify DNA of other 10 types of pathogens, including Neisseria meningitidis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Rubivirus, mumps virus, adenovirus (type 3), adenovirus (type 7), respiratory syncytial virus B and parainfluenza virus type 2, indicating a good specificity. Furthermore, a dry-reagent assay was developed using air-drying and freeze-drying process. The performance of dried reagents did not change after 10 days storage at 50 ℃, meaning the dried reagents could be stored at room temperature (25 ℃) for more than six months. The dry-reagent LAMP assay also successfully amplified MTB DNA from several clinical samples within 20 min. In conclusion, the developed LAMP assay together with isothermal amplifier could rapidly detection MTB.
Sujets)
Humains , Mycobacterium tuberculosis/génétique , Indicateurs et réactifs , Sensibilité et spécificité , Techniques d'amplification d'acides nucléiques/méthodes , ADNRésumé
A molecular diagnostic assay which could be stored at room temperature was developed to rapidly detect Mycobacterium tuberculosis (MTB) based on loop-mediated isothermal amplification (LAMP) technology and dry-reagent process. LAMP uses 4 or 6 primers and Bst DNA polymerase to amplify DNA at a constant temperature. The results showed that the LAMP assay could detect the amplification of IS6110 target gene within 20 min using real-time fluorescence signal detection. The sensitive of LAMP assay was similar to the PCR technology while the precision of PCR was better than LAMP (coefficient of variation, LAMP 18.9%, PCR 3.4%), meaning LAMP was more suitable for qualitative detection. The LAMP assay did not amplify DNA of other 10 types of pathogens, including Neisseria meningitidis, Haemophilus influenzae, Staphylococcus aureus, Streptococcus pneumoniae, Rubivirus, mumps virus, adenovirus (type 3), adenovirus (type 7), respiratory syncytial virus B and parainfluenza virus type 2, indicating a good specificity. Furthermore, a dry-reagent assay was developed using air-drying and freeze-drying process. The performance of dried reagents did not change after 10 days storage at 50 ℃, meaning the dried reagents could be stored at room temperature (25 ℃) for more than six months. The dry-reagent LAMP assay also successfully amplified MTB DNA from several clinical samples within 20 min. In conclusion, the developed LAMP assay together with isothermal amplifier could rapidly detection MTB.
Sujets)
Humains , Mycobacterium tuberculosis/génétique , Indicateurs et réactifs , Sensibilité et spécificité , Techniques d'amplification d'acides nucléiques/méthodes , ADNRésumé
<p><b>OBJECTIVE</b>To investigate the actual condition of the sexual physiological and psychological development of the high school students in Yibin in order to get a reliable basis for sexual education of the teenagers.</p><p><b>METHODS</b>With a proportion of 1% to the whole, 2,770 students were randomly selected from eight high schools in the urban and rural areas of the city. We devised a questionnaire and asked each student to fill in his or her answers presently.</p><p><b>RESULTS</b>So far as sexual physiological and psychological development was concerned, the high school students of Yibin were found rather precocious, with very little sexual knowledge and psychological endurance in sexual affairs and a relative lack of sexual education.</p><p><b>CONCLUSION</b>It is imperative to extend the scope of puberty sexual education in high schools. Teenagers must be taught different kinds of sexual knowledge at different periods of growth as well as how to avoid sexually transmitted diseases and gestation. The sexual knowledge level of the teachers must also be raised. It is a must to establish service or specialist consultation hot lines about sexual knowledge for teenagers. Parents are expected to change their traditional views and assume an active role in the joint efforts of sexual education for their children.</p>