Gustafsdottir Sigrun M, Nordengrahn Ann, Fredriksson Simon, Wallgren Per, Rivera Esteban, Schallmeiner Edith, Merza Malik, Landegren Ulf
The Beijer Laboratory, Department of Genetics and Pathology, Rudbeck Laboratory, Uppsala, Sweden.
Clin Chem. 2006 Jun;52(6):1152-60. doi: 10.1373/clinchem.2005.065847.
Nucleic acid amplification allows the detection of single infectious agents. Protein-based assays, although they provide information on ongoing infections, have substantially less detection sensitivity.
We used proximity ligation reactions to detect proteins on bacteria and virus particles via nucleic acid amplification. Antibodies recognizing viral or bacterial surface proteins were equipped with DNA strands that could be joined by ligation when several antibodies were bound in proximity to surface proteins of individual infectious agents.
Detection sensitivities similar to those of nucleic acid-based detection reactions were achieved directly in infected samples for a parvovirus and an intracellular bacterium.
This method enables detection of ligated DNA strands with good sensitivity by real-time PCR and could be of value for early diagnosis of infectious disease and in biodefense.
核酸扩增可检测单个感染因子。基于蛋白质的检测方法虽然能提供有关正在进行的感染的信息，但其检测灵敏度要低得多。
我们利用邻近连接反应，通过核酸扩增来检测细菌和病毒颗粒上的蛋白质。识别病毒或细菌表面蛋白的抗体配备有DNA链，当多个抗体靠近单个感染因子的表面蛋白结合时，这些DNA链可通过连接反应连接在一起。
对于细小病毒和一种细胞内细菌，在感染样本中直接实现了与基于核酸的检测反应相似的检测灵敏度。
该方法能够通过实时PCR以良好的灵敏度检测连接的DNA链，在传染病的早期诊断和生物防御方面可能具有价值。
