2. INTRODUCTION
MITOCHONDRIAL GENOME
Mitochondrias are organelles
ubiquitous to the cytoplasm of all
eukaryotic cells of animals, higher
plants, and some microorganisms.
Mitochondria generates energy
for cellular processes by
producing ATP through oxidative
phosphorylation.
Although most DNA is packaged
in chromosomes within the
nucleus, mitochondria also have a
small amount of its own DNA.
This genetic material is known as
mtDNA.
2
Cyclospora Cayetanensis
Is an intestinal coccidian
protozoan parasite. It has a
worldwide distribution, and
transmission is through the
fecal-oral route or by
ingestion of contaminated
water or food.
This parasite infects the
gastrointestinal tract of
returning travelers , causing
acute diarrheal disease.
3. 3
INTRODUCTION
As an eukaryotic parasite, C. cayetanensis has a
mitochondrial genome. As mitochondrial genomes are
evolving more quickly than nuclear genomes , molecular
typing assays developed based on mitochondrial
sequences generally have high resolution.
The mtDNA of C. cayetanensis has about 500 copies per
cell; molecular diagnostic tools targeting mitochondrial
sequences may have the additional advantage of higher
detection sensitivity than those targeting nuclear genes,
making the mitochondrial genome a potential marker for
the development of a genotyping tool .
5. 5
MATERIALES Y MÉTODOS
FUNDAMENTO
• Técnica de amplificación enzimática In vitro donde se
obtienen millones de copias de un fragmento específico
de ADN.
• Genotipos en el estudio: CM 1 CM 9
APLICACIÓN
• qPCR amplifica y cuantifica simultáneamente el producto del
DNA replicado.
• Durante la investigación, la qPCR, fue utilizada para hacer la
cuantificación del genoma en las diferentes muestras y para
analizar la cantidad de expresión existente.
qPCR
10. DISCUSSION
10
“In a previous study, eight single-nucleotide variants
(SNVs) and one 7-bp multiple-nucleotide variant
(MNV) in the mitochondrial genomes were detected
between two C. cayetanensis isolates from China and
the USA, making the mitochondrial genome a
potential marker for the development of a
genotyping tool”.
AUTHOR CONTRIBUTION OR
“In theory, the qPCR developed in this study could be
used for rapid assessment of genetic heterogeneity in C.
cayetanensis among specimens in events of augmented
number of cyclosporiasis cases. Previously, a MLST assay
targeting five loci with short tandem repeats (STRs) was
developed for C. cayetanensis”.
Guo Y, et al.
“Although large cyclosporiasis outbreaks have
occurred almost annually in North America and some
European countries during the past 20 years, the
prevention and control of cyclosporiasis outbreaks
are still a major public health challenge”.
Hedberg CW, et al.
Tang K, et al.
11. 11
CONCLUSSIONS
The availability of
Cyclospora
cayetanensis genome
sequence provides
opportunities for
research and
development of
multiple other
diagnostic tools.
The evaluation of genetic
polymorphisms in the same
parasitic species, caused by
geographical variability, can
detect different behaviors
of the same pathogen, which
is useful to focus a more
specific treatment taking
into account the genomic
versatility.
This parasite infects the gastrointestinal tract of returning travelers and immunocompromised patients, causing acute diarrheal disease.
Cyclospora infection may be self-limiting, so antimicrobial therapy is not required in every case
It has a worldwide distribution, and transmission is through the fecal-oral route or by ingestion of contaminated water or food not likely to be transmitted person-to-person.
Histologic changes in mucosal biopsies include mild villous blunting, and surface epithelial disarray.
Infection causes a flulike illness with nausea, vomiting, weight loss, and explosive watery diarrhea lasting 1 to 3 weeks.
Infection may be asymptomatic, and Cyclospora can infect immunocompromised or immunocompetent patients.
Cyclospora is found worldwide. Most reports of cyclosporiasis in wealthy countries involve residents of or travelers returning from resource-limited countries
PCR: FUNDAMENTO- PARA QUE
FUNDAMENTO PCR: AMPLIFICACIÓN ENZIMÁTICA DE UN FRAGMENTO ESPECIFICO DE ADN, ES ENTE CASO, GENOTIPOS.PCRQ PARA HACER CUANTIFICACIÓN DEL GENOMA Y SABER LA CANTIDAD DE EXPRESIÓN QUE HABÍA
Descripcion de lo que estaban analizando . 9 genotipos de 5 continentes ( China, Perú, Nepal, Indonesia, Guatemala, Spain)
RESULTADOS DE AMPLIFICACIÓN:
Figura a.
en los resultados de la amplificación, todos arrojaron bandas por debajo de 500 pB, lo que indica que los genes aislados son de bajo peso molecular
Además, Al comparar los diferentes genotipos de las continentes analizadas, se puede apreciar que los de China y Perú 21453 y 21473, respectivamente, son similares, y en comparación con el 21462 de china se observan dos genes notablemente marcados.
Figua B.
En las cepas peruanas se da la expresión de dos genes por encima de 1000 pB
Existen variaciones de nucleótidos múltiples en el genotipo 21473 de Perú , los demás son constantes con una secuencia de GTTATTA. ( Perú es diferente respecto a España, china, Guatemala y Nepal).
Abajo se encuentra un polimorfismo de un solo nucleótido en las dos cepas de China
1. To improve the investigation of cyclosporiasis outbreaks, a high-resolution multilocus sequence typing (MLST) tool has been developed recently for C. cayetanensis based on whole genome sequence data. -molecular typing assays developed based on mitochondrial sequences generally have high resolution. In a previous study, eight single-nucleotide variants (SNVs) and one 7-bp multiple-nucleotide variant (MNV) in the mitochondrial genomes were detected between two C. cayetanensis isolates from China and the USA, making the mitochondrial genome a potential marker for the development of a genotyping tool.
