Recently, the State Key Laboratory of Respiratory Disease and the Zhang Tianyu Research Team from Guangdong-Hong Kong-Macau Joint Laboratory for Infectious Diseases have made breakthroughs in the research direction of Klebsiella pneumoniae (Kp): the successful construction of a marker-free autoluminescent Klebsiella Pneumoniae (SfAlKp), which can not only establish a high-throughput in vitro drug screening platform, but also realizes real-time, rapid and efficient detection of the anti-Kp activity of drugs in living mice. It only takes 2-5 hours from infection, treatment to preliminary judgment of drug activity in vivo.

For the first time, the team used the endogenous Xer-dif recombination enzyme system and Tn7 transposon system to insert the sequence of the lux genes into the genome of Kp, to successfully construct SfAlKp. Advantages of SfAlKp include:

1. High luminous intensity: In the darkroom, ordinary household cameras can clearly capture the blue and green light emitted by the SfAlKp in the flat panel. For the first time in the world, the dual-weak light technology is used to simultaneously capture the glowing SfAlKp in living mice and the clear outline of the mice.

2. Strong luminescence stability: All single colonies randomly picked after multiple passages can emit strong light.

3. The insertion of the sequence of lux genes did not affect the drug sensitivity and the growth status of Kp. The use of SfAlKp can achieve rapid, real-time and continuous drug screening in vitro and activity evaluation in vivo, which falsely shortens the screening time, reduces the consumption of manpower and material resources, and improves the screening efficiency of antimicrobial drugs with good prospects for clinical applications.

The research results have recently been published in Biosensors and Bioelectronics in District 1 of the Chinese Academy of Sciences. The co-first authors of this paper are Tian Qianrong from the University of Chinese Academy of Sciences and Anhui University, and Gao Yamin from the University of Chinese Academy of Sciences. This project has received strong support from the cooperating units Guangzhou Chest Hospital and Guangzhou Medical University, funded by the National Science and Technology Major Special Project in Major Drug Development, the Chinese Academy of Sciences Project, and the Guangdong Special Support Program.

Original paper : https://linkinghub.elsevier.com/retrieve/pii/S0956-5663(20)30904-0