Technical improvement to prevent DNA degradation of Leptospira spp. in pulsed field gel electrophoresis.
Leptospirosis is a public health problem. Infection with pathogenic Leptospira occurs by exposure to many environments and is traditionally associated with occupational risk activities. Pulsed-field gel electrophoresis was used to investigate the epidemiological relatedness among Leptospira isolates. However, analysis by PFGE yielded inconclusive data as a result of extensive DNA degradation. This degradation can be significantly reduced by the inclusion of thiourea in the electrophoresis buffer, improving the analysis of DNA banding patterns.
Ribeiro
R L
RL
WHO/PAHO Collaborating Center and National Reference Laboratory for Leptospirosis, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, Brazil. rribeiro@ioc.fiocruz.br
Machry
L
L
Brazil
J M V
JM
Ramos
T M V
TM
Avelar
K E S
KE
Pereira
M M
MM
D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods; D016172:DNA Fingerprinting / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000037:antagonists & inhibitors*; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000235:genetics*; D007922:Leptospirosis / Q000382:microbiology; D013890:Thiourea / Q000494:pharmacology*
D004269:DNA, Bacterial; D003851:Deoxyribonucleases; D013890:Thiourea