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Article in Chinese | WPRIM | ID: wpr-236337


<p><b>OBJECTIVE</b>Eplets mismatch based on HLAMatchmaker software evaluates the clinical application of kidney transplantation.</p><p><b>METHODS</b>In 239 cases of renal transplant,merits of methods of the traditional HLA six antigen matcheing criteria, cross reaction groups standard and Eplets mismatch based on HLAMatchmaker standard were compared respectively.</p><p><b>RESULTS</b>The number of mismatchs with three methods in 239 cases, were grouped according to low-high mismatchs. The results revealed that HLAMatchmaker algorithm could significantly increase the number of low mismatchs group 54 (22.6%), compared with the HIA group 19(7.9%) and CREGs group 32 (13.4%). The comparison was discovered statistical significance among the three groups (P<0.001), so the comparison between each group was.</p><p><b>CONCLUSION</b>HLAMachmaker of donor-recipients matching, is a more efficient, time-saving and high sensitivity matching solution to allograft renal transplantation.</p>

Adolescent , Adult , Aged , Female , Histocompatibility Testing , Methods , Humans , Kidney Transplantation , Male , Middle Aged , Software , Transplantation, Homologous , Young Adult
Chinese Journal of Virology ; (6): 410-414, 2013.
Article in Chinese | WPRIM | ID: wpr-339936


To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.

Adolescent , Adult , Aged , Conserved Sequence , Cytomegalovirus , Genetics , Cytomegalovirus Infections , Diagnosis , Virology , DNA, Viral , Blood , Female , Humans , Immunosuppressive Agents , Blood , Kidney Transplantation , Male , Middle Aged , Polyomavirus , Genetics , Polyomavirus Infections , Diagnosis , Virology , Real-Time Polymerase Chain Reaction , Methods , Reproducibility of Results , Sensitivity and Specificity , Species Specificity , Tacrolimus , Blood , Time Factors , Tumor Virus Infections , Diagnosis , Virology , Viral Load , Young Adult
Article in Chinese | WPRIM | ID: wpr-235330


<p><b>OBJECTIVE</b>To investigate the application of ABO blood group genotyping in complicated ABO blood group type.</p><p><b>METHODS</b>Ten specimens of complicated ABO blood group were genotyped by sequence specific primer PCR (PCR -SSP), and confirmed by DNA sequencing and alignment. Six hundred and ten blood samples typed by ABO immunoassay were as control of genotyping.</p><p><b>RESULTS</b>Ten cases of complicated blood type were identified by high resolution PCR- SSP as rare ABO blood groups: cis-AB01 (3 cases), B(A)04 (2 cases), cisAB02, B(A)02, Bel03, Bw12 and Ael05, confirmed by DNA sequencing. Genotyping and serotype detected 610 cases ABO blood group were coincident, and the frequency of A, B, AB and O were as 28.69%, 27.54%, 8.2% and 35.57% respectively. According to the genotypes, the highest frequency subgroup was O1 (32.87%), the lowest was A2 (0.66%).</p><p><b>CONCLUSION</b>PCR -SSP could type the ABO blood group accurately, but also the sub-group of blood type. However, special designed high resolution PCR -SSP or DNA sequencing is needed to identify the complicated blood groups.</p>

ABO Blood-Group System , Genetics , Blood Grouping and Crossmatching , Methods , Genotype , Humans , Polymerase Chain Reaction , Sequence Analysis, DNA
Article in Chinese | WPRIM | ID: wpr-254532


<p><b>OBJECTIVE</b>To investigate polymorphisms of killer cell immunoglobulin-like receptor gene (KIR) in renal transplant recipients from southern Zhejiang.</p><p><b>METHODS</b>KIR genotypes were analyzed by PCR-SSP in 416 renal transplant recipients, and the genotype frequencies were compared with populations from Eastern China and worldwide.</p><p><b>RESULTS</b>All 16 known KIR genes were detected in the renal transplant recipients, and KIR2DL4, 3DL2-3, 3PD1 were found in all. As a pseudogene, 2DP1 has a high genotype frequency (99%). The frequencies of KIR2DL1, 2DL3, 3DL1, 2DS4 have ranged from 92.1% to 98.8%. Compared with 11 groups in Eastern China and other countries, the KIR2DL2 phenotype frequency was higher (34.6%) than those of Shanghai, Zhejiang and Jiangsu populations (P<0.05). Among 41 genotypes, three have not been reported previously. The most common genotype was AA1, with a frequency of 43.51%, which was significantly lower than those of Jiangsu and Northern Zhejiang.</p><p><b>CONCLUSION</b>Renal transplant recipients from southern Zhejiang share similar features with Eastern China Han population with regard to KIR polymorphisms, but also have unique frequencies for KIR genotypes.</p>

Adolescent , Adult , Aged , China , Female , Gene Frequency , Genotype , Humans , Kidney Transplantation , Methods , Male , Middle Aged , Polymorphism, Genetic , Receptors, KIR , Genetics , Young Adult