RESUMO
Antibody-mediated rejection (AMR) induced by donor-specific anti-HLA antibodies (DSA) remains a major cause of long-term graft loss after kidney transplantation. Currently, the presence of DSA cannot always be determined at a specific allele level, because existing donor HLA typing is low resolution and often incomplete, lacking HLA-DP, and occasionally HLA-C and HLA-DQ information and historical donor DNA samples are not available for HLA retyping. Here we present a novel, non-invasive technique for obtaining donor DNA from selectively expanded donor cells from urine of renal transplant recipients. Urine-derived cells were successfully expanded ex vivo from 31 of 32 enrolled renal transplant recipients, and with DNA obtained from these cells, donor HLA typing was unambiguously determined for HLA-A, -B, -C, -DRB1, -DQA1, -DQB1, -DPA1 and -DPB1 loci by next-generation sequencing. Our results showed 100% concordance of HLA typing data between donor peripheral blood and recipient urine-derived cells. In comparison, HLA typing showed that DNA derived from urine sediments mainly contained recipient-derived DNA. We also present the successful application of our novel technique in a clinical case of AMR in a renal transplant recipient. Urine-derived donor cells can be isolated from kidney transplant recipients and serve as a suitable source of donor material for reliable high-resolution HLA genotyping. Thus, this approach can aid the assessment of DSA specificity to support the diagnosis of AMR as well as the evaluation of treatment efficacy in kidney transplant recipients when complete donor HLA information and donor DNA are unavailable.
Assuntos
Transplante de Rim , Alelos , Genótipo , Rejeição de Enxerto/genética , Antígenos HLA/genética , Teste de Histocompatibilidade , Humanos , Doadores de Tecidos , TransplantadosRESUMO
Rotavirus was detected in 52% of 2328 stool specimens collected from children with acute gastroenteritis admitted to three sentinel hospitals in Mainland China from January 2006 to December 2007. G3P[8] (42%) was the most common strain. Despite being common globally, only 18 (2%) G9-positive samples were identified. The VP7, VP4, VP6, and NSP4 genes were sequences for 13 of the G9 strains with G9P[8] being most common and showing the same origin as G9 strains reported in other countries. One G9P[6] strain was possibly derived by reassortment between earlier Chinese G9 strains and more recent local P[6] strains.
Assuntos
Diarreia/epidemiologia , Gastroenterite/epidemiologia , Infecções por Rotavirus/epidemiologia , Rotavirus/genética , Pré-Escolar , China/epidemiologia , Diarreia/virologia , Gastroenterite/virologia , Genes Virais , Genótipo , Humanos , Lactente , Epidemiologia Molecular , Prevalência , RNA Viral/genética , Vigilância de Evento Sentinela , Análise de Sequência de RNARESUMO
OBJECTIVE: Building a method which can examines virus pathogenic in gastroenteritis excrement specimen. METHODS: Choosing six positive specimens which tested in our laboratory, include adenovirus, calicivirus, rotavirus, bocavirus, astrovirus and enterovirus. Through sequence-independent single primer amplification(SISPA) constructs a gene bank. Looks up the viral gene fragment in gene bank. RESULTS: Obtaining corresponding viral acid sequence in six specimens. CONCLUSION: This research can examine enterovirus and the virus which cause diarrhea, It make a foundation for further studies the viral cause of disease which the examination not yet discovered at present.
