High-speed gel microelectrophoresis, a new and easy approach for detection of PCR-amplified microbial DNA from environmental and clinical samples in microgels using conventional equipment.
OBJECTIVE
Microelectrophoresis allows the detection of DNA bands using minimal amounts of sample in a short time, but commonly requires the use of special equipment which is not available in all laboratories. This fact has limited the application of this technique in microbiology despite its advantages. In this work, we describe a new approach to perform gel microelectrophoresis, named high-speed gel microelectrophoresis (HSGME), and its application for rapid detection of bacteria, protozoa and viruses in clinical, vegetal and environmental samples.


RESULTS
Aliquots of 0.4-1 microl of PCR product were loaded in 2 cm 1% agarose microgels and electrophoresed at high voltage (125 V cm(-1)) in conventional submarine horizontal mini-slabs. By using HSGME, single-DNA bands obtained after specific-PCR useful in diagnosis of different diseases caused by micro-organisms were detected in 5 min.


CONCLUSIONS
HSGME is a rapid and easy procedure applicable to detection of microbial genes, which is carried out using conventional equipment and thus can be performed in any research and diagnostic laboratory.


CONCLUSIONS
The performance of HSGME saves up to 90% time, material and energy costs, as well as laboratory hazardous wastes including carcinogenic agents used for visualizing DNA bands.
Velázquez
E
E
Departamento de Microbiología y Genética, Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, Spain. evp@usal.es
Vega
Y
Y
Trujillo
M E
ME
Peix
A
A
Sancho
P
P
Valverde
I
I
Tejedor
C
C
Chordi
A
A
Nájera
R
R
Velázquez
E
E
D019883:Alkyl and Aryl Transferases; D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001826:Body Fluids / Q000382:microbiology / Q000469:parasitology / Q000821:virology; D016785:Cryptosporidium parvum / Q000235:genetics / Q000302:isolation & purification; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004777:Environment; D056890:Eukaryota / Q000235:genetics / Q000302:isolation & purification*; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008904:Miniaturization; D016133:Polymerase Chain Reaction; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000235:genetics; D012231:Rhizobium / Q000235:genetics / Q000302:isolation & purification; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
D012337:RNA, Ribosomal, 18S; D004247:DNA; D019883:Alkyl and Aryl Transferases; C024274:adenylate isopentenyltransferase; D012194:RNA-Directed DNA Polymerase