ABSTRACT
Objective To investigate the diagnostic value of the immediate early antigen(IE) mRNA by nucleic acid sequence-based amplification(NASBA) in peripheral blood cytomegalovirus (CMV) infection,and to establish and promote the diagnosis method for CMV.Methods Five to seven ml blood was taken from 32 patients at 3 week and 7 week after renal transplantation to detect serum cytomegalovirus antigen and antibody expression by NASBA,Real time-PCR and enzyme-linked immunosorbent assay (ELISA) sensitivity and specificity were compared.Results The results of CMV detection in 32 renal transplanted patients respectively showed that the positive rate of peripheral blood IE-mRNA by NASBA was 45.8％ (15/32) ;The positive rate of HCMV-DNA in blood by Real time-PCR was 45.8％ (15/32).Using ELISA,the positive rate of HCMV-(IgG +IgM) was 37.5％ (14/32).IE-mRNA and HCMV-DNA had higher sensitivity and specificity and lower false positive rate than HCMV-(IgG +IgM).The positive rates of IE-mRNA by NASBA,Real time-PCR and ELISA were 92.8％,71.5％ and 42.8％ respectively in the 14 cases.Conclusion The nucleic acid amplification method (NASBA based sequence) and Real time-PCR are sensitive,rapid diagnosis methods of HCMV infection,with higher sensitivity and specificity and lower false positive rate than traditional ELISA.And NASBA detection of IE-mRNA has good value for auxiliary clinical diagnosis.
ABSTRACT
ObjectiveTo investigate the relation of the human cytomegalovirus pp67-mRNA with the active infection in renal transplant recipients and the role in the direction for antiviral therapy.Methods Total 128 samples were collected from peripheral blood treated with EDTA of 32 recipients at the 3rd week or the 7th week after transplantation.IFA was used to detect CMV pp65 antigen,and NASBA was used to detect pp67 mRNA assay.Compared with CMV-Ag,the recipients were followed up to observe the change of pp67 in the cases of the active infection and the CMV disease.ResultsCompared with the proportion between two groups,44 samples out of 128 samples showed CMV-Ag pp65 positive,while pp67-mRNA were positive in 21 samples.8 patients had been found positive CMV-Ag pp65 and pp67-mRNA; 12 recipients were clinically diagnosed with HCMV disease.In the early stage of the HCMV infection,the sensitivity and specificity of the CMV-Ag pp65 ( 91.7％,50.0％ ) was higher than that of pp67-mRNA ( 66.7％,95.0％ ).During the periods of symptoms appeared and the late stage of clinical treatment,the specificity of pp67-mRNA was higher than that of CMV-Ag pp65,while the sensitivity sit on the contrary.And in the antiviral therapy course,the indicator of antiviral therapy pp67 turned negative more quickly than the CMVAg,the therapy course could be reduced.ConclusionsBoth of CMV-Ag pp65 and of pp67-mRNA have clinical significance.Testing for CMV-Ag pp65 is suitable for detecting HCMV active infection at the early stage,which was important for in time anti-virus treatment in the clinical settings.The test for pp67-mRNA is a quick method to accquire the accurate results; therefore,it can be used as the reference criteria for the clinical anti-virus treatment.
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Objective To investigate the expression of complement split product C4d and its significance in renal allograf tissue of chronic rejection in rats.Methods The healthy closed population Wister rats and SD rats were used as donator and acceptor in renal transplantation.The chronic rejection model of renal transplant in rats was established and the rats were divided into 2 groups.The rats in experimental group were given Mycophenolate mofetil(MMF)(10 mg/kg) and those of the control group were given nothing except CsA(5 mg?kg~(-1)?d~(-1)) for 10 days.At the 12th week of renal transplantation,the allograft was tested by light microscope,and the pathological changes of renal grafts and the expression of C4d in peritubular capillaries were observed.Results On the 12th week of renal transplantation,the morphology changes of chronic rejection was observed in the experimental group and obvious C4d deposition was detected in peritubular capillaries of renal allograft tissue,with significant difference compared with those of the control group(all P