[The first case of rabies in Sweden in 26 years. Inform travellers abroad about risks and treatment following suspected infection]. / Första rabiesfallet i Sverige på 26 år. Informera utlandsresenärer om risker och behandling efter misstänkt smitta.

In June 2000, a case of rabies was diagnosed in Stockholm. The patient, a 19-year-old woman, had been bitten by a dog in Thailand three months earlier. She was admitted with a 2-day history of pain and paresthesia at the exposure site (right arm), along with anxiety. Her neurological symptoms progressed, and during the following week she developed the typical signs of furious rabies. Despite intensive care, her condition deteriorated continuously, and she died 18 days after onset of symptoms. The diagnosis was not considered until five days after admission to the hospital. A saliva sample was obtained and the diagnosis confirmed by virus isolation in mouse neuroblastoma cells. Although Sweden is free of rabies, the diagnosis should be considered in patients with encephalitis after having visited a rabies endemic area. Tourists must be informed of the vital importance of post-exposure prophylaxis after suspected infection.

Seroprevalence of viral childhood infections in Eritrea.

BACKGROUND: The seroprevalence of viral childhood infections in Africa has not been thoroughly investigated. The relatively recently discovered human parvovirus B19 (B19) and human herpesvirus 6 (HHV-6) have received particularly little attention. OBJECTIVE: To investigate the seroprevalence of viral childhood infections in different Eritrean populations and to define groups at high risk for infection. STUDY DESIGN: Five population groups in Eritrea have been examined to define the prevalence of specific antibodies to several childhood viruses. The study population of more than 400 persons consisted of children, pregnant women, female sex workers and members of a secluded tribe called Rashaida. RESULTS: All groups showed a high prevalence of antibodies to measles and HHV-6 (> 85%). For rubella, the seroprevalence was very high in all adult groups (93-99%) except the Rashaida group (71%). The mumps prevalence was surprisingly low in the Rashaida group (29%) compared to 46-85% in the other adults. Late encounter of mumps and rubella was also observed among the Rashaidas. The pattern of antibodies to B19 showed a higher seroprevalence in all groups (56-91%) compared to what has been reported from the western world. CONCLUSION: The findings represent what might be expected in an unvaccinated population. The exception was the Rashaidas, which had low seroprevalences and late encounter of mumps and rubella. This is of importance because it makes this tribe vulnerable to these infections, which are associated with complications when acquired in adult age. Also noteworthy is the high frequency of antibodies to HHV-6 and particularly B19 in all groups, indicative of an early encounter of both these viruses.

Seroprevalence of human herpes virus 8 in different Eritrean population groups.

BACKGROUND: Human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma. In the US and Europe, HHV-8 is believed to be mainly sexually transmitted, but reports from some African countries suggest non-sexual transmission. OBJECTIVES: To find out more about HHV-8 seroprevalence and transmission in Eastern Africa. STUDY DESIGN: In this study, 411 serum samples from different population groups in Eritrea (children, pregnant women, female sex workers and members of the isolated Rashaida tribe) were examined for HHV-8 antibodies with an immunofluorescence assay detecting antibodies to latent and lytic HHV-8 antigens. RESULTS: Antibodies to HHV-8 latent antigen were found in 0-2% of Eritrean children, 5% of pregnant women, 8% of female sex workers and 26% of Rashaidas, respectively. No correlation was found between detectable HHV-8 antibodies and seropositivity to HIV or herpes simplex 2. CONCLUSIONS: These results suggest that HHV-8 infection is relatively common in Eritrea and that viral transmission occurs predominantly through non-sexual route in this region.

BK virus as the cause of meningoencephalitis, retinitis and nephritis in a patient with AIDS.

BACKGROUND: The two widely spread human polyomaviruses, BK virus (BKV) and JC virus (JCV) establish latency in the urinary tract, and can be reactivated in AIDS. JCV might cause progressive multifocal leucoencephalopathy, but although up to 60% of AIDS patients excrete BKV in the urine there have been few reports of BKV-related renal and/or neurological disease in AIDS. OBJECTIVE: To report on an AIDS patient with progressive renal and neurological symptoms involving the retina. DESIGN: Case report. SETTING: Venhälsan, Söder Hospital, Stockholm, Sweden. METHODS: The brain, eye tissue, cerebrospinal fluid, urine and peripheral blood mononuclear cells were analysed by nested PCR for polyoma-virus DNA. Macroscopical and microscopical examination were performed of the kidney and brain post mortem. Immunohistochemical stainings for the two BKV proteins, the VP1 and the agnoprotein, were performed on autopsy material and virus infected tissue culture cells. RESULTS: BKV could be demonstrated in the brain, cerebrospinal fluid, eye tissues, kidneys and peripheral blood mononuclear cells. CONCLUSION: During 6 years, approximately 400 cerebrospinal fluid samples from immunosuppressed individuals with neurological symptoms have been investigated by PCR for the presence of polyomaviruses. BKV DNA has, so far, only been found in the case reported here. Although reports of BKV infections in the nervous system are rare, there is now evidence for its occurrence in immunocompromised patients and the diagnosis should be considered in such patients with neurological symptoms and signs of renal disease. The diagnosis is simple to verify and is important to establish.

Prevalence of herpes simplex virus types 1 and 2, cytomegalovirus, and varicella-zoster virus infections in Eritrea.

BACKGROUND: Herpesviruses establish latent infections in their hosts for life. The scarcity of data that exists in regard to herpesvirus infections in many African regions, could partly be due to the mild nature of their primary infections and the lack of means for their proper diagnosis. However, in recent decades the alarming spread of HIV infection in Africa and associated frequent reactivation of herpesvirus infections is leaving less room for neglect. This seroprevalence study is intended to help in the evaluation of the prevalence of herpesvirus infections in Eritrea. OBJECTIVE: To evaluate the spread of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), cytomegalovirus (CMV), and varicella-zoster virus (VZV) infections. STUDY DESIGN: The study population groups comprise female sex workers (FSW), former guerrilla fighters, truck drivers, port workers, a tribe called Rashaida, pregnant women, children under 5 years of age, and children over 5 years of age. The groups of pregnant women and children under and over 5 years of age were included to form a background for the evaluation of groups considered at risk for sexually transmitted or blood borne infections. RESULTS: All study groups had a high seroprevalence of HSV-1 infections ( > 80%), except for the children under 5 years of age. The FSW had the highest prevalence of HSV-2 infections, 80%, followed by guerrilla fighters, truck drivers, port workers, pregnant women, children, and the Rashaidas. Positivity for antibodies against CMV was > 90% in all studied populations. The prevalence of VZV infections was surprisingly low in the tribe of Rashaida, 44% compared to more than 90% in the other adult groups tested for VZV (P = 0.0001). CONCLUSION: The study shows that the prevalence of HSV-2 in the risk group of FSW was high, which could partly be explained by their sexual behaviours. HSV-2 was particularly low in the Rashaida group and, as expected, in the children. The low prevalence of VZV observed in the Rashaida is of importance since it makes them vulnerable to infection with varicella during their inevitable integration with the other tribes in their society.

Identification of BK virus in a patient with acquired immune deficiency syndrome and bilateral atypical retinitis.

OBJECTIVE: To report the clinical and histopathologic characteristics of BK virus (BKV) retinitis. DESIGN: Case report. TESTING: The clinical features of bilateral retinitis in a 29-year-old homosexual white male with the acquired immune deficiency syndrome (AIDS) included focal, mottled fundus pigmentation, and haloes, as documented by fundus photography. After death of the patient, the left eye was studied by light microscopic and immunohistochemical examination. The nested polymerase chain reaction (PCR) was used to detect viral deoxyribonucleic acid (DNA) in the right eye and other nonocular tissues. The specificity was then confirmed by restriction enzyme analysis. RESULTS: The retina of the left eye showed focal necrosis and contained cells with intranuclear staining for the BKV VP1 protein. In the right eye, BKV DNA was detected in the retina and other tissues by nested PCR. Autopsy showed that BKV infection was also present in the brain, kidneys, and peripheral blood mononuclear cells. CONCLUSIONS: A number of pathogens may cause retinitis in patients with AIDS. The authors have shown that BKV should be included among those pathogens and that some clinical features may suggest the presence of BKV retinitis.

[A review of the current research on prions. The evidence suggests the possibility of transmission of the mad cow disease to humans]. / Genomgång av den aktuella forskningen kring prioner. Fakta talar för att galna ko-sjukan kan överföras till människor.

Further evidence of the transmissibility of bovine spongiform encephalopathy (BSE) across the species barrier from cow to man has been derived from epidemiological analysis and the characterisation of prion strains. Recent research has shown the persistence of prions after experimental transmission to resistant murine species, and subclinical persistence in cows. The accumulation of pathological prion proteins in tonsils and appendix has been demonstrated prior to clinical confirmation of the presence of the new variant of Creutzfeldt-Jakob disease. Current prion research is focused on the involvement of B lymphocytes as carriers, on the species barrier and cellular receptors, and on macromolecules involved in the conformational change from normal to pathological prion proteins.

Development of human antibodies against linear antigenic and immunogenic regions of respiratory syncytial virus (RSV) nucleocapsid and phospho-proteins shows the site-directed characteristics.

BACKGROUND: The successful development of an RSV vaccine requires a better understanding of the pathogenesis of primary infection, susceptibility to reinfection, and the immunopathology of enhanced illness in children immunized with a non-replicating RSV candidate vaccine. The exact role of different immune parameters in RSV pathogenesis remains controversial. OBJECTIVES: To study the contribution of antibodies directed to the linear antigenic and immunogenic regions of the N and P proteins in the titer rise and avidity maturation of total anti-RSV antibodies. STUDY DESIGN: The occurrence of antibodies directed against three linear antigenic and immunogenic regions in each of the nucleocapsid (N): N3 (Thr11 to Gly30), N25 (Ser231 to Ala250) and N39 (Thr371 to Leu391), and the phospho-(P) proteins of respiratory syncytial virus (RSV), subgroup A: P49 (Pro91 to Asp110), P56 (Ser161 to Lys180) and P62 (Glu221 to Phe241), were analyzed in ELISA with (a) 32 paired sera from humans with recent or previous RSV subgroup A and/or B infection diagnosed by conventional ELISA, detection of antigen in nasopharyngeal aspirates and measurement of antibody avidity change; and (b) 40 RSV antibody-positive sera (HCS) obtained from patients during their convalescence from RSV infection and possessing clearly demonstrable titers of RSV IgG in conventional enzyme immunoassays (EIA) based on whole RSV antigen. RESULTS: The titer rise of antibodies directed to the combined three peptides representing the RSV N protein was well correlated with the rise in anti-RSV antibodies measured in whole antigen ELISA. Surprisingly, the rise in antibodies against a truncated main C-terminal epitope (Gln381 to Leu391) of the N protein (represented by subgroup A specific sequence of the N39/1 peptide) was inversely correlated with the titer rise of total anti-RSV antibodies. The titer rise of antibodies to the C-terminal linear site of the RSV N (N39/1) protein was subgroup-specific during the course of primary RSV infection. A titer rise in antibodies to the C-terminal linear sites of RSV N (i.e. N39/1) and P (P62) proteins had a dominating appearance in sera from newborn infants (6-7 months) and from patients with RSV reinfections. Anti-RSV antibody titers of late convalescent sera correlated with the titers of antibodies directed to the C-terminal linear site of RSV P (P62) protein. The avidity maturation of the anti-RSV immune response followed the titer rise of anti-P62 antibodies during the course of primary or secondary RSV infection.

Prevalence of infection with HIV, hepatitis B and C viruses, in four high risk groups in Eritrea.

BACKGROUND: Little is known about the prevalence of infections in different population groups in Africa, and about the influence of living conditions on the spread of infections. This study is the first of its kind in the state of Eritrea and is expected to serve as an evaluation of the situation in the country. OBJECTIVE: A serosurvey for human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) was carried out during the summer of 1995 in Massawa, a small sea port in Eritrea (East Africa) in four groups considered to be at risk for these infections. STUDY DESIGN: The study subjects were former Guerrilla Fighters, Female Sex Workers, Truck Drivers, and Port Workers. Participants from a tribe called Rashaida were believed to be at low risk, and thus served as a control. RESULTS: The Female Sex Workers had the highest incidence of HIV-1 infection, 29%, compared to 10% for Port Workers, and 3% for Guerrilla Fighters. On the other hand presence of HBsAg, indicating a high prevalence of hepatitis B carrier status, was highest in the Guerrilla Fighters, followed by the Rashaidas, and lowest in the Female Sex Workers. The Female Sex Workers were further tested for antibodies against HBV and the results revealed that 53% of them, 5%, had antibodies against HBcoreAg. Excluding the possibility of an acute infection at sampling time, three of them became HBsAg carriers. Surprisingly, our group of Truck Drivers did not show HIV-1 infection, and no HIV-2 infections were seen in the whole cohort. CONCLUSION: The study shows that the described groups have different prevalences of infection with HIV, hepatitis B and C which can partly be explained by their living conditions.

Polymerase chain reaction for detection of JC virus DNA in cerebrospinal fluid: a quality control study. European Union Concerted Action on Viral Meningitis and Encephalitis.

The sensitivity and specificity of PCR of CSF for the diagnosis of progressive multifocal leuko encephalopathy is estimated at 75 and 98.5%, respectively. However, inter-laboratory and inter-technique variations have been shown to produce wide variations. A 10-fold dilution series of JC virus in cerebrospinal fluid was prepared and circulated for 'blind' evaluation in laboratories participating in a European Union Concerted Action on Virus Meningitis and Encephalitis. Six of seven laboratories returned results with sensitivity of between 10 and 1 JCV DNA copy equivalents per 10 microl of CSF, one laboratory detected 10(5) copies per 10 microl of CSF. These results demonstrate the feasibility of using virus diluted in CSF for comparison of PCR techniques, and that the range of sensitivity of JCV PCR in proficient laboratories is between 10 and 1 copy equivalents per 10 microl of CSF.

Analysis of PCR as a tool for detection of JC virus DNA in cerebrospinal fluid for diagnosis of progressive multifocal leukoencephalopathy.

Two polyomaviruses, JC virus (JCV) and BK virus (BKV), affect humans. JCV is the causative agent of progressive multifocal leukoencephalopathy (PML), and detection of JCV in the central nervous system (CNS) is a prerequisite for confirmation of the disease. BKV is generally not associated with neurological disease, but involvement of BKV in patients with CNS disorders has been reported. In the present study polyomavirus DNA was detected by a nested PCR at a sensitivity corresponding to the detection of 10 copies of JCV DNA in 10 microliters of cerebrospinal fluid (CSF). CSF samples from 212 patients with neurological symptoms and immunodeficiencies were investigated for the presence of polyomavirus DNA. Of 128 human immunodeficiency virus (HIV)-infected patients, 14 (11%) had JCV DNA in their CSF, and all 14 patients had clinical PML. BKV DNA was detected in one HIV-infected patient with neurological symptoms not compatible with PML. Among 84 HIV-negative patients, 6 (7%) had detectable JCV DNA in their CSF, confirming PML in patients with clinical conditions of T-cell lymphoma, chronic lymphatic leukemia, status postliver transplantation, congenital immunodeficiency, sarcoidosis, and immunodeficiency of unknown origin. The specificity of the PCR was confirmed by a clinical follow-up study which showed full agreement between the detection of JCV DNA in CSF and clinically manifest PML. The described PCR is a rapid, reproducible, and easily performed assay.

Detection of respiratory viruses in nasopharyngeal secretions with immunofluorescence technique for multiplex screening-an evaluation of the Chemicon assay.

BACKGROUND: Antigen detection with immunofluorescence is an efficient method for diagnosis of respiratory tract infections, but has previously not allowed for simple screening of many respiratory viruses. Pools of monoclonal antibodies against various respiratory viruses are now available, and are potentially important tools for improvement of antigen detection in nasopharyngeal samples. OBJECTIVE: To evaluate the commercially available Chemicon immunofluorescence assay (IFA; respiratory viruses panel and identification kit), an indirect IFA containing a pool of monoclonal antibodies for screening for influenza A, B, respiratory syncytial virus (RSV), parainfluenza 1, 2, 3 and adenovirus, and the respective individual antibodies. STUDY DESIGN: Ninety-six frozen preparations from nasopharyngeal secretions or bronchoalveolar lavages were retrospectively examined with the assay, and the results compared with other IFAs for antigen detection and cell culture isolation obtained in the everyday routine. Nasopharyngeal preparations from 300 children with lower respiratory tract infections at Beijing Children's Hospital during the 1994-1995 winter season were also examined. RESULTS: The sensitivity of the Chemicon assay compared to the combined results of routine IFA and isolation was 89% and specificity 92%. If five identifications of RSV made with the Chemicon assay alone were regarded to be truly positive, the specificity was 100%. A viral etiology was identified in 105/280 (38%) evaluable samples drawn from the Chinese children (influenza A 20%, RSV 14%, adenovirus 3% and parainfluenza 1, 2 or 3, 7%). CONCLUSION: One problem with the Chemicon assay was that for around 4-13% of samples there was a non-specific staining in the screening assay, necessitating stainings for verification. Despite this, the assay is an excellent tool for identification of viral respiratory tract infections, giving an increased sensitivity compared to direct immunofluorescence assays.

Viral diagnosis by antigen detection techniques.

BACKGROUND: Diagnosis of viral infections can be obtained in the early stages of a disease by detection of viral antigens directly in the clinical specimen. This has become an important tool for rapid virus diagnosis. METHODS: Antigens produced during virus infections can be detected either in cells collected from the site of infection by immunohistological investigation or in secretions and blood by solid phase immunoassays (IA). Viruses causing acute respiratory infections can be diagnosed in cells from the respiratory tract, viruses causing vesicular eruptions in epithelial cells from skin scrapings, rabies virus in nerve cells of the brain or epithelial cells from skin and cornea and cytomegalovirus (CMV) matrix antigen, pp65, can be detected in peripheral blood leukocytes (PBL) by immunofluorescence (IF) or immunoperoxidase techniques. The quality of specimens can be easily checked during the reading of results. Some IAs for antigen detection, such as detection of HBsAg and HIV p24 antigen in blood are standardized and sensitive. Others give less sensitive results because of the variation of quality of the clinical specimen. The latex agglutination tests are mainly used for rapid detection of virus or viral antigens in faeces: rota-and adenoviruses; the method may not be very sensitive but yields a result within a few minutes. Assays detecting viral nucleic acids are more sensitive than antigen detection tests because of a tremendous amplification of gene segments obtained by the polymerase chain reaction (PCR). So far such assays are time consuming and expensive and are mainly used in specific clinical situations. RESULTS: After introduction of specific monoclonal antibodies (Mabs), the antigen detection techniques are increasingly used. the need for quality control, trained staff, and standardized reagents and methods for specimen collection and preparation is now being appreciated. IF for viral respiratory viruses is used for diagnosis and epidemiological studies all over the world. Likewise, IF is still the method most often used for rabies diagnosis. For CMV, the pp65 matrix antigen is shown to be a sensitive marker closely correlated with clinical symptoms. Its detection by the IF technique has proven to be superior to other techniques for prediction of CMV pneumonia in bone marrow transplant patients. IAs are currently used in fully automated systems for large scale diagnosis based on antigen detection in serum specimens. Increase of antibody specificity on the solid phase by use of Mabs directed against the most abundant viral antigen in the clinical specimen shortens the reaction time; this has been employed in most of the constantly appearing new rapid diagnosis kits based on the immunoassay principle. CONCLUSION: Although, in virology, more sensitive results are obtained by the gene detection method, PCR, directly in clinical samples, viral antigen detection tests are, after the introduction of Mabs for diagnostic purposes, increasingly used because of their low demand on laboratory equipment, their rapid and early result and relatively low cost. Antigen detection is successfully used directly in clinical specimens for rapid diagnosis of many viral infections as well as for identification of tissue culture isolated viruses. With Mab-based IAs the reaction time is shortened and new rapid, almost 'instant test' kits are appearing on the market.

No association of JC virus with Alzheimer's disease or astrocytomas.

OBJECTIVES: To investigate if JC virus (JCV) can be involved in the pathogenesis of Alzheimer's disease (AD) and astrocytomas. STUDY DESIGN: A nested polymerase chain reaction (PCR) was used for the detection of JCV DNA in autopsy brain material (cerebral white matter) and cerebrospinal fluid (CSF) specimens from patients with AD and age-matched control patients without neurological diseases, together with biopsies from patients with astrocytomas (grades 3 and 4). Brain autopsy material from AIDS patients with progressive multifocal leukoencephalopathy (PML) was examined as positive control material. RESULTS: JCV DNA was detected by PCR in only one of the 17 brain autopsies from patients with AD, but in none of the 26 control patients without neurological diseases and in none of the 5 astrocytoma biopsies. JCV DNA was, however, detected in the brain material from two patients with PML. CONCLUSION: Our results show that JCV infection does not seem to be directly involved in the pathology of AD or in the development of astrocytomas. In addition, since no viral DNA was detected in CSF specimens from 43 patients without PML (17 with AD and 26 elderly controls), our results suggest that the finding of JCV DNA in CSF correlates to PML.

Association of serum antibodies against defined epitopes of human papillomavirus L1, E2, and E7 antigens and of HPV DNA with incident cervical cancer.

In order to provide a large-scale evaluation of the association with cervical cancer of antibodies against human papillomavirus (HPV) antigens, sera from 233 patients with primary, untreated cervical cancer and from 157 healthy age- and sex-matched blood donors were analyzed for IgG and IgA antibodies against HPV-derived peptide antigens and against bovine papillomavirus. Several serological responses were strongly associated with cervical cancer, notably the IgG response against the HPV 16 epitopes L1:13 (Relative risk [RR]: 5.3), E2:9 (RR: 2.9), and E7:5 (RR: 4.3), and the IgA response against an HPV 18 E2-derived antigen (245:18, RR: 3.1). HPV DNA in corresponding cervical tumors was analyzed by Southern blotting (SB) and polymerase chain reaction (PCR) in 47 patients. Sixty-six percent of the patients carried HPV DNA as determined by SB, 91% of patients analyzed by PCR. Neither the antibody responses, nor the presence of HPV DNA were significantly associated with the biological properties of the tumors.

Nested PCR for detection of BK virus and JC virus DNA.

BACKGROUND: A nested polymerase chain reaction (PCR) was developed to detect BK virus (BKV) and JC virus (JCV) DNA sequences. The unique clevage site for BamHI restriction enzyme was located in the JCV amplimer and cleavage was used to differentiate between BKV and JCV. STUDY DESIGN: Twenty-three urine specimens from 17 bone marrow recipients with haemorrhagic cystitis and one liver transplant patient were tested for the presence of BKV and JCV DNA. Four brain tissue specimens (paraffin embedded brain tissues and a fresh frozen brain biopsy) and 5 cerebrospinal fluids from 3 AIDS patients and one liver transplant patient, all with progressive multifocal leukoencephalopathy (PML), were also examined by PCR. RESULTS: The sensitivity of the PCR was 10 genomes for each virus. BKV DNA was detected in 15 urine specimens from 12 bone marrow transplant patients. JCV DNA was detected in 4 cerebrospinal fluids and 4 brain tissues from patients with PML. CONCLUSION: Our results show that the nested PCR is a sensitive and rapid assay that can be used for diagnosis of BKV and JCV infections. The cerebrospinal fluid appears to be a suitable material for diagnosis of JC virus reactivation in the brain.

Viral pathogens and clinical manifestations associated with acute lower respiratory tract infections in children of the Sudan.

BACKGROUND: Incomplete knowledge regarding the viral agents causing respiratory infections in children living in developing countries impedes diagnosis and management of patients. OBJECTIVES: To assess the role of viral pathogens in Sudanese children presenting with acute lower respiratory tract infections (ALRI). STUDY DESIGN: The study population consisted of patients presenting with ALRI at the Children's Emergency Hospital in Khartoum during 2 periods (December 1987 to April 1988 and September 1990 to March 1991). Identification of viral infections was based an antigen detection by immunofluorescence and enzyme-linked immunosorbent assay (ELISA) on nasopharyngeal secretions and/or serology. RESULTS: After exclusion of children with measles, 102 and 111 children, respectively, were prospectively enrolled in the study during the 2 periods. Their ages ranged between one mouth and 14 years (mean 2.0 years). Radiologic pulmonary infiltrations were detected in 135 (66%) of the 206 patients who had chest radiographs, whereas 7 (3%) showed lobar pneumonia. The case fatality rate was 2.3%. Of 83 virus infections detected, 79 were in children < years and consisted mainly of respiratory syncytial virus (RSV, 28%), followed by parainfluenza (7%), adenovirus (5%) and influenza A (2%). Infiltrates on radiographs were significantly less often found in virus-infected cases than in ALRI-cases with negative virus tests. CONCLUSIONS: RSV predominantly infected young infants (

Nested PCR for detection of BK virus and JC virus DNA.

BACKGROUND: A nested polymerase chain reaction (PCR) was developed to detect BK virus (BKV) and JC virus (JCV) DNA sequences. The unique clevage site for BamHI restriction enzyme was located in teh JCV amplimer and cleavage was used to differentiate between BKV and JCV. STUDY DESIGN: Twenty-three urine specimens from 17 bone marrow recipients with haemorrhagic cystitis and one liver transplant patient were tested for the presence of BKV and JCV DNA. Four brain tissue specimens (paraffin embedded brain tissues and a fresh frozen brain biopsy) and 5 cerebrospinal fluids from 3 AIDS patients and one liver transplant patient, all with progressive multifocal leukoencephalopathy (PML), were also examined by PCR. RESULTS: The sensitivity of the PCR was 10 genomes for each virus. BKV DNA was detected in 15 urine specimens from 12 bone marrow transplant patients. JCV DNA was detected in 4 cerebrospinal fluids and 4 brain tissues from patients with PML. CONCLUSION: Our results show that the nested PCR is a sensitive and rapid assay that can be used for diagnosis of BKV and JCV infections.