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1.
Leg Med (Tokyo) ; 14(1): 36-9, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22182593

RESUMO

The DNA extraction performance of three low-throughput extraction systems was evaluated. The instruments and respective chemistries all use a similar extraction methodology that involves binding DNA to a coated magnetic resin in the presence of chaotropic salt, washing of the resin to remove undesirable compounds, and elution of DNA from the particles in a low-salt solution. The AutoMate Express™ (Life Technologies Corporation, Carlsbad, CA), EZ1® Advanced XL (Qiagen Inc., Valencia, CA), and Maxwell® 16 (Promega Corporation, Madison, WI) were compared using a variety of samples including: blood on swabs, blood on denim, blood on cotton, blood mixed with inhibitors (a mixture of indigo, hematin, humic acid, and urban dust) on cotton, blood on FTA® paper, saliva residue on cigarette butt paper, epithelial cells on cotton swabs, neat semen on cotton, hair roots, bones, and teeth. Each instrument had a recommended pre-processing protocol for each sample type, and these protocols were followed strictly to reduce user bias. All extractions were performed in triplicate for each sample type. The three instruments were compared on the basis of quantity of DNA recovered (as determined by real-time PCR), relative level of inhibitors present in the extract (shown as shifts in the C(T) value for the internal PCR control in the real-time PCR assay), STR peak heights, use of consumables not included in the extraction kits, ease of use, and application flexibility. All three systems performed well; however extraction efficiency varied by sample type and with the preprocessing protocol applied to the various samples.


Assuntos
Impressões Digitais de DNA/instrumentação , DNA/isolamento & purificação , Alelos , Sangue , Osso e Ossos/química , Células Epiteliais/química , Cabelo/química , Humanos , Masculino , Papel , Reação em Cadeia da Polimerase , Saliva/química , Sêmen/química , Têxteis , Dente/química
2.
Emerg Infect Dis ; 16(3): 441-6, 2010 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-20202419

RESUMO

Data regarding the type, frequency, and distribution of tick-borne pathogens and bacterial agents are not widely available for many tick species that parasitize persons in the southern United States. We therefore analyzed the frequency and identity of pathogens and bacterial agents in ticks removed from humans and subsequently submitted to the Texas Department of State Health Services, Zoonosis Control Program, from October 1, 2004, through September 30, 2008. The data showed associations of bacterial agents and potential vectors. Tick-related illnesses may pose unidentified health risks in areas such as Texas, where incidence of human disease related to tick bites is low but well above zero and where ticks are not routinely suspected as the cause of disease. Cause, treatment, and prevention strategies can be better addressed through collecting sufficient data to establish baseline assessments of risk.


Assuntos
Borrelia/isolamento & purificação , Ehrlichia/isolamento & purificação , Rickettsia/isolamento & purificação , Infestações por Carrapato/parasitologia , Doenças Transmitidas por Carrapatos/microbiologia , Carrapatos/microbiologia , Animais , Vetores Aracnídeos/microbiologia , Borrelia/classificação , Borrelia/genética , Ehrlichia/classificação , Ehrlichia/genética , Humanos , Rickettsia/classificação , Rickettsia/genética , Especificidade da Espécie , Texas , Doenças Transmitidas por Carrapatos/transmissão , Carrapatos/classificação
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