Detection of Human Cytomegalovirus (HCMV) and Porcine Endogenous Retrovirus (PERV) with One Step Extraction Method / 감염과화학요법
Infection and Chemotherapy
;
: 12-16, 2010.
Artigo
em Inglês
| WPRIM
| ID: wpr-225196
ABSTRACT
BACKGROUND:
Xenotransplantation is thought to be one of the alternative methods to overcome the shortage of human organs for transplantation. Recipients should be immunosuppressed for graft survival, and thus, there is a need for developing diagnostic modality that can detect diverse infections originating from animals and recipients rapidly, in the early stage, and with high sensitivity using small volume of samples. This study was carried out to develop a fast, simple, and robust technique for the preparation of HCMV DNA and PERV RNA using small volume of samples. MATERIALS ANDMETHODS:
Nucleic acids were extracted from serially diluted samples with one step extraction method as well as with Qiagen kit. The presence of genomic DNA of human cytomegalovirus (HCMV) and porcine endogenous retrovirus (PERV) was detected by PCR and specific primer set, respectively. RNA of HCMV and PERV was extracted and then detected by RT-PCR and specific primer set, respectively. For absolute quantification of HCMV, standard curve was established by real time PCR.RESULTS:
HCMV DNA and PERV RNA were prepared from culture supernatant and cells for PCR or RT-PCR with one step extraction method. It was possible to extract both the DNA and RNA from the samples in about 20 minutes with one step extraction method in a single tube. HCMV and PERV could also be detected by PCR and one step extraction method, respectively. It was also good with small quantity samples.CONCLUSIONS:
One step extraction method is simpler and faster method than other extraction methods when there are two types of DNA and RNA viruses in one sample. From these results, we could see that the one step extraction method could be very useful in detecting HCMV and PERV rapidly from the pig cells or organ transplanted recipients with a small amount of sample.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Vírus de RNA
/
Transplante Heterólogo
/
DNA
/
RNA
/
Ácidos Nucleicos
/
Reação em Cadeia da Polimerase
/
Transplantes
/
Retrovirus Endógenos
/
Citomegalovirus
/
Sobrevivência de Enxerto
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
/
Humanos
Idioma:
Inglês
Revista:
Infection and Chemotherapy
Ano de publicação:
2010
Tipo de documento:
Artigo
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