Tompkins L S, Krajden M
Diagn Microbiol Infect Dis. 1986 Mar;4(3 Suppl):71S-78S. doi: 10.1016/s0732-8893(86)80044-x.
A previous study of the detection of Campylobacter jejuni in fecal samples spotted directly onto nitrocellulose filters before hybridization revealed a relatively low sensitivity and some false-positive results. We have investigated two factors that interfere with the detection of Campylobacter jejuni in fecal samples: interfering substances that create false-positive background signals and nonspecificity of the probe. Heterologous deoxyribonucleic acid probes bound nonspecifically to partially extracted fecal samples, indicating that the major basis for false-positive background was not due to homologous sequences. The problem of false-positive results could be reduced, but not eliminated, by extensive deoxyribonucleic acid extraction procedures applied to the clinical sample. The relative concentration of protein in each sample may be an important contributor to nonspecific binding. Other measures, such as sonication, glass bead fragmentation, and column separation, were not helpful. Development of a species-specific deoxyribonucleic acid probe derived from sequences encoding 16S ribonucleic acid is underway.
先前一项对空肠弯曲菌的检测研究发现，将粪便样本直接点样在硝酸纤维素滤膜上然后进行杂交，其灵敏度相对较低且存在一些假阳性结果。我们研究了两个干扰粪便样本中空肠弯曲菌检测的因素：产生假阳性背景信号的干扰物质以及探针的非特异性。异源脱氧核糖核酸探针与部分提取的粪便样本非特异性结合，这表明假阳性背景的主要原因并非同源序列。通过对临床样本采用广泛的脱氧核糖核酸提取程序，假阳性结果的问题可以得到减少，但无法消除。每个样本中蛋白质的相对浓度可能是导致非特异性结合的一个重要因素。其他措施，如超声处理、玻璃珠破碎和柱分离，并无帮助。目前正在研发一种源自编码16S核糖核酸序列的种特异性脱氧核糖核酸探针。
