[Application of M-RAPD technique to obtain the genomic fingerprints of various pathogenic microbials].
OBJECTIVE
To obtain genomic fingerprints of different pathogenic microbials and make certain whether their patterns can be used in the identification of microbials by means of multiplex random amplified polymorphic DNA analysis (M-RAPD).


METHODS
Arbitrary primers of 10 oligonucleotides were randomly grouped, and various microbials chromosomal DNA were amplified with three combinatorial primers at a special higher annealing temperature. The products were detected by 15 g/L agarose electrophoresis and the patterns were analyzed by the software of Gelworks 1d Intermediate.


RESULTS
Specific and resistant DNA fingerprints for different pathogenic microbials with combinatorial primers were gained. The profiles were clear, well-distributed and the number was great. The products of the three primers included most products of every two primers and would appear with no relation to their length, but small products had more opportunity; three primers could provide information contents half as many again as that two primers could provide for the same pathogenic microbials. There were obvious differences among different drug-resistant strains and between the drug-resistant strains and the corresponding reference strains, but different strains of the same microorganism had more similarity than discrepancy. The analytic data of the software of Gelworks 1d Intermediate also support our results.


CONCLUSIONS
M-RAPD is a simple and rapid technique for the identification of different kinds of pathogenic microbials, and it can provide rich genetic information.
Laboratory of Biochemistry and Molecular Biology, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Gang
G
Liu
Chuan-Min
CM
Tao
Chao-Yang
CY
Zhai
D016172:DNA Fingerprinting; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis; D004926:Escherichia coli / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D005819:Genetic Markers; D014644:Genetic Variation; D005838:Genotype; D019105:Random Amplified Polymorphic DNA Technique; D013211:Staphylococcus aureus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D013296:Streptococcus pneumoniae / Q000145:classification / Q000235:genetics / Q000302:isolation & purification
D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers