ABSTRACT
Objective:To evaluate the specificity and sensitivity of ABI 7000,7300 and 7500 real-time PCR instruments.Methods:Random,double-blind clinical trial;We selected 30 patients with HBsAg and HBV DNA positive for positive group and 30 health volunteers with HBsAg and HBV DNA negative for negative group.We also selected three of HBV DNA positive and negative samples for repeat test.Results:The 30 samples of positive group were detected HBV DNA.The 30 samples of negative group were not detected HBV DNA.However,in positive group,the HBV DNA levels were different.The related index of ABI 7000 with ABI 7300,ABI 7500 and ABI 7300 with ABI 7500 was 0.89,0.89 and 0.98,respectively.The HBV DNA level was highest detected with ABI 7300 instrument,and then with ABI 7000 instrument.The lowest level was detected with ABI 7500 instrument.There was significance among these instruments(p
ABSTRACT
<p><b>OBJECTIVE</b>To study the quantity of anti-R7V in individuals infected with HIV and AIDS patients and its relation with the progression of disease.</p><p><b>METHODS</b>ELISA and precipitation and other methods were used to investigate the quantity of anti-R7V in asymptomatic long-term survivors and AIDS patients.</p><p><b>RESULTS</b>Positive rate and quantity of anti-R7V were higher in the HIV active ones and AIDS. It showed that the quantity and positive rate of anti-R7V were rather high in dissolving test.</p><p><b>CONCLUSIONS</b>It is strong suggestion for anti-R7V to obstruct the replication of virus by interfering the connection between HIV with CCR5 or CXCR4 and so it impossible HIV entering to CD4+ T cells.</p>
Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Acquired Immunodeficiency Syndrome , Allergy and Immunology , Follow-Up Studies , HIV Antibodies , Blood , HIV Infections , Allergy and Immunology , HIV Long-Term Survivors , HIV-1 , Allergy and Immunology , Receptors, CCR5 , Physiology , Receptors, CXCR4 , PhysiologyABSTRACT
<p><b>BACKGROUND</b>To construct recombinant vector expressing antisense RNA to CCR5 in eukaryotic cells and obtain recombinant pseudovirus, which will be used to block HIV-1 infection.</p><p><b>METHODS</b>The DNA fragment targeted against the initional part of CCR 5 mRNA translation was amplified by using RT-PCR from peripheral blood mononuclear cells (PBMCs) and cloned into retroviral vector pLXSN, then transfected into packaging cell (PA317) with lipofectAMINE. After 2-3 weeks selecting with G418, the pseudovirion in the survival cell's supernatant was detected with RT-PCR (FQ),then was used to infect NIH/3T3 cell.</p><p><b>RESULTS</b>The psuedovirion packed from expression vector of sense/antisense RNA to CCR5 had infected NIH/3T3 cell successfully. The vector had incorporated into its genome and transcripted into RNA.</p><p><b>CONCLUSIONS</b>The gene fragment of antisense RNA to CCR5 could be obtained from PBMCs and transfected into eukaryotic cell with retroviral vector. The results made a great foundation for studying its inhibiting effect on HIV-1 infection.</p>
Subject(s)
Animals , Mice , 3T3 Cells , Eukaryotic Cells , Metabolism , Gene Expression , Genetic Vectors , Plasmids , Genetics , RNA, Antisense , Genetics , Receptors, CCR5 , Genetics , TransfectionABSTRACT
<p><b>BACKGROUND</b>To search a novel sensitive, specific and lower cost method applicable for quantitative analysis of the hepatitis B virus DNA extensively.</p><p><b>METHODS</b>Quantitative analysis of the DNA from 100 sera by real-time PCR with Sybr green 1. The results of Sybr's assay were compared with the results obtained with Taqman's fluorescent quantitative assay.</p><p><b>RESULTS</b>Taqman real-time PCR could help evaluate the level of virus reliably. The results of Sybr's assay were in agreement with the Taqman's assay, but detection rate was lower.</p><p><b>CONCLUSIONS</b>Sybr green 1 real-time PCR appeared to be convenient and cheap, but detection rate was lower.</p>