1. Rapid detection of duck hepatitis A virus
genotype C using reverse transcription
loop-mediated isothermal amplification
(February 2014)
Chuanfeng Li, Zongyan Chen, Chunchun
Meng, Guangqing Liu

2. INTRODUCTION

3. INTRODUCTION

4. HEPATITIS
Hepatitis
is
an
inflammatory disease that
affects the liver, this is
generated
infectious
,
Immune or toxic reaction .
It is also considered,
depending on its etiology,
a sexually transmitted
disease.

5. HEPATITIS A
Hepatitis A is a viral liver disease that can
cause mild to severe symptoms
The hepatitis A virus is transmitted by
ingestion of contaminated food or
beverages or by direct contact with a
person infected with the virus, Is
associated with lack of clean water and
poor sanitation
.

6. RT-PCR
•Amplification of RNA through previous
synthesis of DNA copy, using a reverse
transcriptase, followed by several cycles of
conventional PCR, this technique can
determine the expression of genes in
different tissues

7. OBJECTIVE
Evaluate
the
specificity
and
sensitivity in the conserved regions
of 3D gene of duck hepatitis A virus
genotype C (DHAV – C), using
reverse transcription loop-mediated
isothermal
amplification
(RTLAMP), for the prevention and rapid
disease diagnosis.

8. MATERIALES Y MÉTODOS

9. MATERIALES Y MÉTODOS

10. MATERIALES Y MÉTODOS

11. MATERIALES Y MÉTODOS

12. MATERIALES Y MÉTODOS
• DISEÑO PRIMER´S
Tabla 1. Detalles de RT-LAMP y RT-PCR primers diseñados para la detección de
DHAV-C GX1201 cepa.

13. MATERIALES Y MÉTODOS
• EXTRACCION DEL RNA Y DNA
• RNA
Temperatura
:-80
Temperatura
:-20 ° C

14. RT-LAMP

15. RT-PCR

16. MATERIALES Y MÉTODOS

17. MATERIALES Y MÉTODOS

18. RESULTADOS
Fig 1. Determinación de las condiciones óptimas para el ensayo de RT -LAMP

19. RESULTADOS
Fig 2. Detección de productos de RT-LAMP por métodos alternativos.

20. RESULTADOS
Fig 3. Especificidad del ensayo de RT-LAMP y RT-PCR.

21. RESULTADOS
Tabla 2. Comparación de ensayo de RT-LAMP y RT-PCR en muestras clínicas.

22. DISCUSSION

23. CONCLUSIONS
• The detection limit of RT-LAMP was about 100 times
higher than of the RT-PCR, which suggests that the RTLAMP was more sensitive than RT-PCR.

24. CONCLUSIONS
• The RT-LAMP is a simple technique of early diagnosis,
rapid, highly specific, sensitive and cost effective for the
detection and monitoring of potential infection DHAV-C.
• Primer design is crucial for the differentiation of DHAV-C
from other genotypes or the specificity of this assay.

25. CONCLUSIONS
• The RT-LAMP can be widely used in the field or
laboratory with limited resources, because their
results can be easily detected with the naked
eye due to increased turbidity or discoloration.

26. Manuela Diaz Valenzuela

27. Juliana Caro Palacio

28. THANKS