REFERENCIAS - F. Luo, Z. Li and G. Dai et al., Simultaneous detection of different bacteria by microchip electrophoresis combined with universal primer-duplex polymerase chain reaction, Journal of Chromatography A, https://doi.org/10.1016/j.chroma.2019. 460734 - Octava edición Biología Molecular, Lina María Martínez Sánchez, Natalia Vargas Grisales, Laura Mejía Cardona, Felipe Osorio Ospina, Sergio Ramirez Pulgarín.

1. Sofía Adarve Rengifo
III semestre
Facultad de Medicina
Universidad Pontificia Bolivariana
2020 – 1

2. INTRODUCTION
PCR (Polymerase Chain Reaction) MCE (Microchip Electrophoresis)
This method amplifies small samples of
genes, DNA or RNA segments that are
present in different mixtures.
It is use (among others) for DNA
manipulation and identification of virus and
bacterias
This technique uses an electric field in order
to separate charge molecules such as
proteins and nucleic acids.
It is widely used because of its fast analysis
speed and its minimum samples
requirements

3. INTRODUCTION

4. OBJECTIVE
This study aimed to simultaneously
detect two kinds of bacteria by
microchip electrophoresis
combined with universal primer-
duplex polymerase chain reaction
Salmonellaenterica
serovarS.Typhimurium
Pseudomonas
aeruginosa

5. MATERIALES Y MÉTODOS
Staphylococcus
aureus
Listeria
monocytogenes
Escherichia coli Salmonella
Typhimurium
CULTIVO
Medio de cultivo → Luria – Bertani (LB) Broth
Temperatura → 37 °C
Tiempo → 20 horas
Pseudomonas
aeruginosa

6. Es el tipo de MCE mas usado por su alta sensibilidad
MCE – LIF
(laser induced fluorescence)
MATERIALES Y MÉTODOS
Se usó para identificar y
diferenciar C1 y C2
Las moléculas se separan en el campo
eléctrico de acuerdo a su carga y peso
molecular
Las moléculas migran al electrodo
opuesto según su carga
Investigación

7. MATERIALES Y MÉTODOS
Primer dupplex
PCR
Amplificación
enzimática in
vitro
A partir de una pequeña muestra
de un segmento de ADN se
obtienen múltiples copias de la
secuencia.
Se necesita:
- Una sola hebra de ADN
- Un primer
- Una DNA polimerasa
Se utilizó para detectar las
concentraciones de P. aeruginosa y
S. Typhimurium por medio de la
cuantificación de C1 y C2
Investigación
Permite amplificar simultáneamente
diferentes secuencias

8. RESULTADOS

9. RESULTADOS

10. DISCUSSION
AUTHOR CONTRIBUTION YES OR NOT
Y. Zhang, et al.
“Under optimal conditions, the detection of S.
Typhimurium by MCE-LIF was achieved within
135 s with a limit of detection (S/N = 3) of
3.37 × 102 CFU mL−1 ”
K.Y. Wang, Y.L. Zeng, X.Y. Yang,
W.B. Li, X.P. Lan
“After centrifugation, the supernatant was
used as a template for the amplification of
bound aptamers by PCR with FITC- or trB-
labeled primers (20–30 cycles of 0.5 min at
94°C, 0.5 min at 52°C, 0.5 min at 72°C, followed
by 5 min at 72°C)."
Y. Zhang, et al.
“The limits of detection for the three bacteria
were ranged from 40 to 70 CFU mL−1.”

11. CONCLUSIONS
Parameters as primer concentration,
MgCl2 concentration and temperature
need to be taken into consideration in
order to develop a sensitive and specific
method of bacteria detection.
The idea of using the method exposed in
this article in the daily medical practice is
tempting, due to the fact that S.
Thymurium is associated with gut diseases
in humans and P. aeruginosa is related
with respiratory problems.
Mixing PCR method with MCE
technique have positive effects in
bacteria determination. When working
together, the LOD (lower limit of
detection) decreases notoriously, which
means that the techniques have
become more sensitive to the colonies
forming units, making the time of
detection shorter.