Arana Escandón, Yasser Isaac Castrillón Cardona, Cristhian Camilo Estudiantes medicina UPB 2017

1. Yasser Isaac Arana Escandón
Cristhian Camilo Castrillón Cardona
Seminario Biología Molecular
2017

2. Introduction
Burkholderia
cepacia
Characteristics
In the Next Slide
Problem
Contamination of medications,
medical devices, and other
health care products.
Scientific article’s reason
An outbreak of
24 patients in
Houston, Texas
Where They discovered: Liquid
Docusate was contaminated
with B.C
In consequence, Domestic
manufacturer had to retire
their product

3. Burkholderia cepacia
Burkholderia:
 They belong to the pseudomonas
 Straight bacilli
 Gramnegative
 They are mobile bacteria with a single flag or with a plume of
polar flagella depending on the species.
 They are mesophilic (15º - 35º Optimum growth temperature)
 Their metabolism is aerobic.

4. Burkholderia cepacia
Burkholderia Cepacia:
 The bacteria attacks patients with Cistic fibrosis
or chronic granilomatous disease.
 Burkholderia cepacia inhabit soil and water and
they can live large periods in humid
environments.

5. Liquid Docusate
The docusate is a laxative that acts by facilitating
the uptake of water by the dregs and its mixture
with the oil substances of the fecal skittle. In this
way softens and lubricates the stool facilitating
the evacuation.

6. Liquid Docusate: relation
with Article
The liquid docusate is the vector by which B.C
generates the outbreak

7. General objective
“To investigate an outbreak of Burkholderia
cepacia complex and describe the measures
that revealed the source.”

8. Métodos
1. Epidemiologia
2. PCR
3. Microbiologia

9. 1. Epidemiología
1. Detección de 3 grupos de pacientes en UCI-
ped. B.C + (Feb 2016) en la misma semana
a) (2 cultivos de esputo y 1 urinario)
*Porque este moo en personas sin FC es
MUY RARO

10. 1. Epidemiología
2. Muestra: Todo paciente que haya estado en
Cuidados intensivos cardiovasculares con B.C +
3. La Investigación buscó factores de riesgo y
posibles relaciones (camas contiguas,
medicamentos, etc).

11. 1. Epidemiología
¿Para qué?
Implementar medidas de prevención con la
muestra
– Lavado de manos
– Aislamiento
– Capacitación a personal de Salud
– Limpieza y desinfección
– Reparación de fugas de agua

12. 1. Epidemiología
4. ¡B.C en heces! → ampliación de
espectro de mecanismos de
transmisión:
 Implementos de aseo
 Alimentos
 Medicamentos
(entéricos-inhalados-tópicos)

13. 2. PCR
• Desarrollada por Kary Mullis en 1.985
• Busca amplificar un fragmento de ADN (clonación
acelular)
1. Desnaturalización del ADN
2. Hibridación de la hebra diana con PRIMER
3. Replicación de la hebra
1.993

14. 2. PCR
• Requiere 20-40 ciclos de 1,5-5 minutos c/u
Etapas:
1. Separación de las hebras a 90°-97°C (ADNp T°estables)
2. Hibridación de hebra de interés con Primer (37-65°C)
3. ADNp elonga los primers (5’→3’) (72-75°C)
*El proceso requiere TERMOCICLADOR
(estandarizó)

15. 2. PCR

16. 3. Microbiología
1. Para los aislamientos se uso rep-PCR (PCR que
usa secuencias extragénicas palindrómicas
como PRIMER)
¿Para qué?
Para evaluar aislamientos (determinar + o – para
B.C)
TÉCNICA
DiversiLab

17. 3. Microbiología
BCSA (Burkholderia Cepacia Selective
Agar)
CULTIVO
Inhibe
Gram +
NO contaminación

18. 3. Microbiología
BCSA (Burkholderia Cepacia Selective
Agar)
CULTIVO
Realizado en:
Agua de grifo
Fregaderos
Productos de uso oral
Medicamentos: Docusato y Albuterol***

19. 3. Microbiología

20. Resultados

25. Discussion

26. Author Approach Yes - Not
Kutty et al As has been recognized in other Burkholderia
species outbreaks associated with oral products,
ventilated patients may be more easily recognized
if exposed given that they are unable to protect
their lower respiratory tracts coupled with the
ease in sampling respiratory specimens during the
course of mechanical ventilation.
Yes
Torbeck et al B. cepacia contaminated products can be
overlooked as a source given the nature of
the organism
Yes*
Torbeck et al & Kim et al This organism is relatively resistant to
preservatives; thus the potential for product
contamination even in the setting of
preservatives exists
Yes
Reik et al Burkholderia cepacia complex is widely
distributed in water, soil, and vegetative matter.
Indeterminate species of B. cepacia complex, such
as in our outbreak, compared to B. cenocepacia
and B. multivorans, are less likely to cause
infections in CF patients, suggesting that it was
unlikely that a CF patient was the origin of the
outbreak
Definitely yes

27. • Basic knowledge of laboratory methods in molecular
biology is a great tool to understand much of areas
such as microbiology and epidemiology
• It is fundamental to develop concepts about the
metabolism and characteristics of microorganisms to
carry out a correct medical practice, making better
clinical and epidemiological approaches
Conclusions

28. Conclusions
• The mechanisms of production and marketing of drugs
that are distributed to health centers should be re
evaluated, and more if this products can be used in a
population inmunosuppresses
• It's necessary that all personal health have knowledge
in preventing the spread of diseases by different
agents, since even though this hasn't been the case,
situations such as these could be frequent in many
health centers

31. “La suerte favorece sólo a la mente preparada”
Louis Pasteur