| Background and Objective: Allogeneic hematopoietic stem cell transplantation, allo-HSCT has been an effective way to treat malignant hematologic disease, aplastic anemia, autoimmune diseases and some solid tumors. Human cytomegalo-virus (hCMV) infection is the early main infection and death-cause to the recipients of allo-HSCT. The prevenient mortality of interstitial pneumonia (I?) caused by CMV infection was above 80 precent. CMV belongs to DNA virus of human herpersviruses, and CMV is very common among people. When infecting normal people, because of the normal immune function of the host, CMV reproduces slow, stays in a latent and concordant state and usually causes no clinical syndrome. When the host immune function especially the cellar immunofunction is suppressed, the latent CMV can be reactivated, and CMV reproduce in a high level constantly, the causes CMV active infection and CMV infectious disease with serious clinical syndromes. At the same time the infectious rate of CMV has been increasing constantly and drawing much attention to by the medical field, then the serious disease caused by CMV infection became a focal point of medical research again. Graft-verse-host disease, GVHD is a main obstacle to the allo-HSCT, and CMV can induce GVHD; affluence the survival of HSCT, delaying the hematopoiesis rebuilding; causing IP. It is a high-dangerous factors of secondary aspergillosis infection, also can be subject to bacteria, fungi andprotozoon infections ,thus lead to to the failure of HSCT. As the occurrence and combined application of many new immune inhibitors, unrelated-HSCT, haploid-HSCT, non-myeloablative-HSCT developed extensively, and HSCT has been developed quickly in recent years. Though the treatments of CMV using ganciclovirus have turned for better, the key of antivirus therapy depends on early-stage diagnose and timely treatment. Using antivirus treatment in time before the occurrence of CMV diseases can prevent and decrease the occurrence of the disease and lower the mortality of HSCT recipients . The early clinical signs and symptoms of the disease by CMV active infection are mostly non-specific, and its early diagnosis mainly depends on laboratorial examination. At present methods for CMV diagnosis are various. Inspite of the more intact diagnostic methods, there is much controversy on which method is simpler, more sensitive and reliable to diagnose CMV early, and then early intervening treatment is in great dilemma. In this paper, we used enzyme-linked immunosorbent assay (ELISA), flow cytometry (FCM), fluorescence quantitative PCR (FQ-PCR/real-time PCR) to examine IgG antibodies and IgM antibodies of CMV, pp65 antigen, quantity of DNA for exploring an effective way to diagnose early CMV infection.Methods and Material: In our research, we used ELISA to detect IgG antibodies, IgM antibodies of CMV; used FCM to detect pp65 antigen; used FQ-PCR to examine HCMV DNA load of blood plasma. Before conditioning we examined CMV IgG, IgM and excluded IgM positive HSCT recipients from our research. We simultaneously examined CMV IgG, IgM, pp65 antigen and DNA load of recipients (the transplantation group) at the day of transfusion of hematopoietic stem cell, and every 10 days after the transfusion of hematopoietic stem cells, of normal HSCT donors (the normal group) before G-CSF mobilization; of leukemic patients (the chemotherapy group)with long term chemotherapy when their WBC to 2.0x109/L after chemo- therapy. We used indirect ELISA: the antigen to examine was in lamellan aperture, washing plate, added Ag-enzyme complex, washing plate again after incubation added substratum, put in darkness to colorate in room temperature,then examined OD valves by enzyme immunoassay (X=450nm); we used indirect simple color method by FCM selecting single-nuclear cells, then fixing, effecting cell membrance, adding pp65 antibodies and mouse IgG1 antigen (the control) then adding rat-anti-mouse IgGl (antibody), then detecting the proportion of positive cells. The FQ-PCR took the outer-marking method: abstrac... |
