ABSTRACT
Objective To establish human metapneumovirus(hMPV)nucleotide rapid,specific and sensitive gene chip detec-tion method,and provide effective diagnostic methods for hMPV detection and diagnosis in shenzhen area.Methods The flu-orescence PCR primer of hMPV was designed for the highly conservative regional gene sequence of hMPV virus.Application of array probe design software designer 4.20 hMPV oligonucleotide detection probe design,oligonucleotide probe sample points to aldehyde slides on the preparation of hMPV gene chip,and parallel compared with conventional reverse tran-scriptase-polymerase chain reaction(RT-PCR),the sensitivity,specificity and repeatability,and was used to evaluate the clinical applicability of the samples.Results hMPV gene chip method to detect hMPV specificity was 96.23%(230/239), the sensitivity was 2.0×101/μl,linear range was 2.0×101~2.0×107/μl,and the repeatability was good.Initial tests in shenzhen area 300 clinical specimens on a nasopharyngeal swab,gene chip method detection rate was 20.3%(61/300),sig-nificantly higher than the conventional RT-PCR method 9.7%(29/300),the difference was statistically significant between the sensitivity of two methods(χ2=39.205,P<0.05).The results were consistent in two methods(kappa=0.360 7).Con-clusion Established hMPV microarray assay,gene chip method to detect hMPV has high specificity and sensitivity,wide linear range and detection.The popularity of hMPV monitoring for laboratory provides a new detecting technology and early diagnosis.
ABSTRACT
<p><b>AIM AND METHODS</b>The properties and sensitivity to acetylcholine of PC12 cells differentiated with nerve growth factor (NGF) have been investigated by using whole-cell clamp technique.</p><p><b>RESULTS</b>When cultured in the presence of NGF, PC12 cells not only differentiated to resemble sympathetic neurons morphologically, but also developed electrical excitability. NGF-treated PC12 cells were highly sensitive to ACh than untreated cells. The I(Ach) proved to be generated by nAChR by pharmacological identification. Nicotinic receptor was characterized by desensitization. The macroscopic I(ACh) was inward rectified and concentration dependent.</p><p><b>CONCLUSION</b>PC12 cells are easily cultured and provides a homogenous population of cells. When culture in NGF, they differentiate to sympathetic-like neurons that contain on their surface neuronal nAChR, it can be used as good model system for studying regulation of a sympathetic neuronal nAChR.</p>