Comparative analysis of a double primer PCR assay with plasma, leukocytes and antigenemia for diagnosis of active human cytomegalovirus infection in bone marrow transplant patients.

The aim of the study was to determine the prognostic value of a double primer PCR assay to detect human cytomegalovirus (HCMV) infection or disease in bone marrow transplant (BMT) recipients. A total of 209 blood samples including peripheral blood mononuclear cells (PBMN), polymorphonuclear (PMN) leukocytes and plasma from 26 BMT recipients were tested by PCR assay. To discriminate between latent and active HCMV infection, 177 blood samples were also tested by a quantitative antigenemia assay. HCMV serology status of donors and recipients was determined before transplantation by an enzyme immunosorbent assay method. Using the double primer PCR assay, the number of positive samples increased by an average of 11.6%. Symptomatic active HCMV infection was diagnosed in 14 (53.8%) out of 26 BMT patients. There was a good association between double primer PCR assay of PMN leukocytes and antigenemia assays for detection of active HCMV infection in all patients. Detection of HCMV DNA in PMN leukocytes of BMT patients by double primer PCR assay can be an alternative method for antigenemia assay. However, quantitative PCR methods will be necessary for monitoring antiviral treatment.

Risk of viral transmission via bone marrow progenitor cells versus umbilical cord blood hematopoietic stem cells in bone marrow transplantation.

Hematopoietic stem cell transplantation (HSCT) is the treatment of choice for children and certain adults with malignant and nonmalignant hematologic disease. Since viral infections are the major problem, this study examined those that might potentially be transmitted to HSCT recipients via bone marrow (BM) versus umbilical cord blood (UCB). BM progenitor cells, peripheral blood leukocytes, and plasma samples were collected from 30 allogenic BM donors. Umbilical cord blood hematopoietic stem cells and plasma samples were also collected from 34 UCB donors. Viral DNA extracted and purified from collected specimens was processed using nested polymerase chain reactions (PCR) to detect human parvovirus B19 (HPV B19), human herpesvirus-6 (HHV-6), varicella-zoster virus (VZV), human cytomegalovirus (HCMV), and Epstein-Barr virus (EBV). The prevalences of HCMV DNA in collected BM progenitor cells versus UCB hematopoietic stem cells were 73% versus 23%, respectively. Conversely, HHV-6 DNA was not detected in any collected specimen by simple PCR. Distribution of the other investigated virus DNAs except EBV DNA was similar in specimens collected from both groups. EBV DNA was not determined in UCB hematopoietic stem cells. The results indicate that the risk of viral transmission to BM transplant recipients via UCB hematopoietic stem cells is less than that with BM progenitor cells.

Hematopoietic stem cell transplantation for beta-thalassemia major: experience in south of Iran.

INTRODUCTION: Allogeneic stem cell transplantation as a curative treatment for thalassemia major was established in Shiraz in 1993. In this article we describe our results of 10 years experience with allogeneic bone marrow transplantation for thalassemia major. METHODS: From June 1993 to January 2003, 112 cases of beta-thalassemia major underwent allogeneic marrow transplantation from HLA-identical or one antigen-mismatched related donors. Conditioning chemotherapy included busulfan (14 to 15 mg/kg), cyclophosphamide (200 mg/kg), and antithymocyte globulin (ATG; 40 mg/kg). Prophylaxis for graft-versus-host disease consisted of cyclosporine, prednisolone, and methotrexate. RESULTS: One hundred twelve patients with a diagnosis of beta-thalassemia major underwent allogeneic marrow transplantation during this period. The mean age of the patients was 9.5 years with the range of 2 to 20 years. The distribution of cases according to the Lucarelli classification were: 27 cases class I, 38 cases class II, and 47 cases class III. Eighty-seven of 112 patients (77.6%) with diagnosis of beta-thalassemia major are living with full engraftment at a median follow-up of 6 years (range 2 to 119 months). CONCLUSION: Allogeneic bone marrow transplantation has changed the outcome of disease dramatically. According to our results stem cell transplantation is the treatment of choice for class I and II (Lucarelli risk groups). Also, we recommend transplantation as a curative method for treatment of class III beta-thalassemic patients.

Qualitative detection of human cytomegalovirus DNA in the plasma of bone marrow transplant recipients: value as a predictor of disease progression.

OBJECTIVE: The aim of this prospective study was to determine whether human cytomegalovirus (HCMV)-DNA detected by polymerase chain reaction (PCR) analysis in the plasma of bone marrow transplant (BMT) patients is a predictor of HCMV disease progression. METHODS: Plasma samples were collected from 15 patients who received allogenic BMTs. Each individual was sampled 1 week before and then weekly for 17 weeks after transplantation. The 270 plasma specimens were processed with a PCR method for detecting HCMV-DNA. Patients were also physically examined for signs or symptoms of HCMV-related disease. RESULTS: Eight (53.5%) of the 15 patients tested positive for HCMV-DNA. Two (25%) of these 8 individuals also had positive PCR findings before transplantation. Six (75%) of the 8 HCMV-DNA-positive patients had positive plasma-PCR results a week before clinical symptoms developed. The other 2 (25%) remained asymptomatic throughout their hospital stay. All 6 symptomatic cases were treated with ganciclovir, and 4 converted to negative plasma-PCR status at a median of 21 days. There was a significant correlation between PCR-detection of HCMVDNA in plasma and presence of HCMV-related symptoms (P < 0.01). CONCLUSION: Qualitative plasma-PCR analysis before and after bone marrow transplantation is a valuable way to screen for HCMV infection in BMT patients. Plasma-PCR monitoring of HCMV activity in this patient group might make it possible to administer an antiviral drug and thus reduce mortality. However, quantitative PCR is still considered the best way to accurately identify active HCMV infection and monitor treatment.

Bone marrow transplantation for hematological disorders--Shiraz experience.

OBJECTIVE: In the past 8 years, 120 cases of hematological disorders were transplanted from the HLA identical donors. METHOD: Using chemotherapy based conditioning regimen with cyclophosphamide 200 mg/kg and busulfan 15-16 mg/kg, 80 cases of beta-thalassemia major and 35 cases of leukemia and five patients with aplastic anemia had received bone marrow transplantation. RESULT: The five-year-survival in thalassemic group was 72%, for leukemic group (acute and chronic) was 58%, and also for aplastic anemia 65%. Transplantation related mortality was the cause of death in 29 cases. The two major causes of death were acute graft versus host disease and poor medical condition of patients before marrow transplantation. CONCLUSION: At the present time, allogenic marrow transplantation is curative mode of treatment for many hematological diseases.

An unusual migration of Taenia hydatigena larvae in a lamb.

The liver and lungs of an four month old, female dead lamb was referred to Veterinary clinic of Shahrekord, Iran by a sheepherder due to outbreak of an unknown disease that caused four deaths in the livestock over a period of one week. Post-mortem examination of the liver showed a massive infection of Taenia hydatigena larvae. Diffuse, spiral and haemorrhagic tracts made by migrating larvae were seen throughout the liver. Large brown to red areas of haemorrhages also appeared on the liver cut surfaces. All the recovered T. hydatigena larvae from migratory canals and hepatic surfaces were all immatures. There was no mature cyst formation. No evidence of pulmonary involvement was found. Histopathological examinations of the liver revealed numerous sections of migratory tracts filled with red blood cells, fibrin and tissue debris. Sections of T. hydatigena larvae were observed at the ends of migratory canals. Hepatocellular degeneration, necrosis, fatty change and infiltration of mixed inflammatory cells, including lymphocytes, plasma cells and macrophages were associated with these tracts. This article reports outbreak of an unusual and severe hepatitis cysticercosa with striking hepatic lesions that caused mortality in a livestock.