Development of molecular markers of Mycobacterium tuberculosis rifampicin resistance gene rpoB by PARMS technology / 生物工程学报

The purpose of this study is to provide a simple and reliable genetic typing approach for molecular drug susceptibility test of Mycobacterium tuberculosis, through the developing of fluorescence molecular marker of rifampicin resistance gene rpoB. Eleven fluorescent molecular markers of the rpoB gene were established by using the sequence difference between the amino acid positions 531, 526, 516, 511 and 513 of rpoB gene of rifampicin-resistant strains and the alleles of rifampicin-sensitive strains, combined with the PARMS technique (Penta-primer amplification refractory mutation system). We used 104 clinical isolates of Mycobacterium tuberculosis to validate this marker and it was verified by sequencing as 100% correct. These samples were also tested with proportional drug sensitivity test. The coincidence rate was 94.23%. The molecular markers had high reliability for genotyping of rpoB gene. It can also detect low-concentration drug-resistant samples (511/533 unit point mutations) whose phenotypic susceptibility cannot be detected. The eleven sets of fluorescent molecular markers could cover 92%-96% of rpoB gene mutation types of rifampicin-resistant strains, and provide new idea for rapid detection of rifampin-resistant Mycobacterium tuberculosis.

Experimental Study of Stress Effects on the Degradation Rate in vitro of Novel Magnesium Alloy Bone Screws / 医用生物力学

Objective To study the effect of stress on the degradation rate in vitro of novel magnesium alloy bone screw. Methods A three-dimensional （3D） model of the tibia fracture was established using the reverse engineering method. Then, based on the FE model, the in vitro degradation experimental device for bone screws was designed. The stress distribution of the screw by finite element calculation was used as the in vitro experimental load, which effectively improved the accuracy and efficiency of the experiment. The experimental samples were divided into four groups. Group A was treated as control group without force application, while Groups B, C and D were subjected to 150, 250 and 350 N axial forces. The influence of different mechanical environment on the degradation rate in vitro of bone screws was investigated. Finally, combining the stress distributions with the degradation experiment results in vitro, the curve between the stress and the degradation rate in vitro of novel magnesium alloy bone screws was obtained. Results Degradation experiments in vitro showed that Group A had the lowest weight loss and hydrogen production, and the average degradation rate was (0.315±0.005) mm/a. While in the stress groups, the weight loss and hydrogen production increased gradually with the axial force increasing. The average degradation rates of Groups B, C and D were (0.379±0.006), (0.469±0.007) and (0.547±0.009) mm/a, respectively. Conclusions When the novel magnesium alloy bone screw was degraded in mechanical environment, the greater stress on the screw would cause the faster degradation rate in vitro. The obtained relationship between the maximum stress and the average degradation rate in vitro of the novel megnesium alloy bone screw provided data support and theoretical guidance for material selection, design and clinical application of magnesium alloy bone screws.