Liu Chunjiang,Huang Junfu,Xia Han,Zhang Xiaoli,Wang Jue,Pan Hong,Fu Weiling.Rapid identification of blood pathogens by capillary electrophoresis-based single strand conformation polymorphism[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(06):537-540.
Liu Chunjiang,Huang Junfu,Xia Han,Zhang Xiaoli,Wang Jue,Pan Hong,Fu Weiling.Rapid identification of blood pathogens by capillary electrophoresis-based single strand conformation polymorphism[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(06):537-540.DOI:
Objective To investigate the feasibility of identifying pathogens in blood infection by capillary electrophoresis-based single strand conformation polymorphism (CESSCP)
and to establish a new method for rapid identification of pathogens. Methods A pair of universal primers was designed and synthesized according to the sequences of 16S rRNA of 7 stains of blood pathogens
including E.coli
S. aureus
P. aeruginosa
S. epidermidis
S. pneumoniae
K. pneumoniae
and E. faecalis. The genomic DNA of above strains was extracted. Then the highly conserved sequences of 7 pathogens were amplified with the universal primers and the extracted DNA as templates. PCR products of 16S rRNA were analyzed by CE-SSCP to discriminate 7 pathogens. Results Our designed universal primers amplified the 16S rRNA of 7 stains of blood pathogens. SSCP electrophoresis showed characteristic peak pattern for the different strains with different peak time respectively
E.coli: 3 999 ms; S. aureus: 4 082 ms; P. aeruginosa: 3 958 ms; S. epidemidis: 4 063 ms; S. pneumoniae: 4 055 ms; K. pneumoniae: 3 980 ms; and E. faecalis: 4 166 ms. The resulting patterns could be used to differentiate all of the pathogens mentioned above. Conclusion Seven pathogens are identified rapidly and conveniently by CE-SSCP.