Uyttendaele M, Schukkink R, van Gemen B, Debevere J
Department of Food Technology and Nutrition, Faculty of Agricultural, UG, Ghent, Belgium.
J Appl Bacteriol. 1994 Dec;77(6):694-701. doi: 10.1111/j.1365-2672.1994.tb02821.x.
NASBAR, an isothermal amplification technique for nucleic acids, was evaluated for the specific identification of Campylobacter jejuni, Camp. coli and Camp. lari. A set of primers and a probe were chosen from the 16S rRNA sequence of Campylobacter. The probe was hybridized in solution with the amplified nucleic acids of 12 Campylobacter species and nine other Gram-negative bacteria. The probe was shown to hybridize specifically to the amplified single-stranded RNA of Camp. jejuni, Camp. coli and Camp. lari in an enzyme-linked gel assay (ELGA). In a Camp. jejuni model system the combination of NASBAR and ELGA was able to detect ca 1000 rRNA molecules. The presence of an excess of Gram-negative bacteria did not influence the sensitivity of detection. A number of 6 cfu of Camp. jejuni, present in a total count of 4 x 10(6) cfu of Gram-negative bacteria, resulted in a positive hybridization signal.
NASBAR是一种用于核酸的等温扩增技术，已针对空肠弯曲菌、结肠弯曲菌和拉氏弯曲菌的特异性鉴定进行了评估。从弯曲菌的16S rRNA序列中选择了一组引物和一个探针。该探针在溶液中与12种弯曲菌属细菌和9种其他革兰氏阴性菌的扩增核酸进行杂交。在酶联凝胶分析（ELGA）中，该探针显示出与空肠弯曲菌、结肠弯曲菌和拉氏弯曲菌的扩增单链RNA特异性杂交。在空肠弯曲菌模型系统中，NASBAR和ELGA的组合能够检测到约1000个rRNA分子。过量革兰氏阴性菌的存在并不影响检测的灵敏度。在革兰氏阴性菌总数为4×10⁶ cfu的情况下，6 cfu的空肠弯曲菌可产生阳性杂交信号。
