bluuebunny/crossref_metadata_2025 huggingface dataset:
Coxsackie virus B4 infection of the mouse pancreas: I. Detection of virus‐specific rna in the pancreas by in situ hybridization
Authors: Vella C. , Easton A. J. , Eglin R. P. , Brown C. L. , Perry L.
Published: March 2007

The pathology of Coxsackie virus B4 (CVB4) infection in a murine model was investigated by in situ hybridization using a biotinylated cDNA probe derived from CVB4. During the acute phase of infection virus RNA sequences were detected in the exocrine pancreas of 60% of mice infected with a pancreotropic variant of CVB4. A positive hybridization signal was observed in other organs in some animals including the heart and liver of 1 mouse 28 days after infection. The cellular distribution of virus RNA sequences corresponded well with the histological findings in most tissues. Possible causes for failure of hybridization in some infected pancreases are discussed in conjunction with potential application of the technique in human pancreas biopsy samples.
Influence of volume expansion on capillary transport in the gut of pichindé virus infected strain 13 guinea pigs
Authors: Katz Murray A. , Starr James F.
Published: March 2007

Hydropenic strain 13 guinea pigs (GP13) injected with Pichindé virus show marked jejunal capillary disruption determined by a reduction in protein reflection coefficient (σ) [Katz and Starr, 1990]. These studies examined whether this is reversed by a 2% Ringer's expansion, and whether anorexia may contribute to reduced σ. GP13 were either infected or given a 60% expected ad lib diet, and were studied while under a 2% Ringer's expansion. The results showed 1) infected GP13 separated into groups with either a persistently low u (volume unresponsive) or a normal σ (volume responsive), but no obvious feature of the animals could predict responsiveness; 2) caloric restriction had minimal to no effect on σ; and 3) both responsive and unresponsive GP had increased heart, lung, and kidney weights. Volume expansion may have salutary effects in some infected GP13, and this is probably due to pressure mediated effects on the capillary wall. The findings support the possibility that conservative management of hemorrhagic fevers may directly ameliorate capillary leak.
Molecular epidemiology of an outbreak of poliomyelitis in South Africa in 1987/1988
Authors: Tsilimigras C. W. A. , Rossouw E. , Schoub B. D.
Published: March 2007

An Outbreak of paralytic poliomyelitis occurred in the Republic of South Africa in 1987/88. The epidemic took place in the Natal/KwaZulu region of South Africa and was due to type 1 poliovirus. Twenty‐four isolates were characterised by 1‐and 2‐ dimensional oligonucleotide mapping and a single strain of wild‐type poliovirus was identified. The same strain of virus was also isolated in other areas of South Africa at the time of the outbreak and has persisted into 1989.
A novel approach to enterovirus typing
Authors: Lee Spencer H. S. , Boulilier James E. , Macdonald Margaret A. , Forward Kevin R.
Published: March 2007

A novel method was developed for typing enteroviruses producing cytopathic effect. Mono‐layers of primary or secondary rhesus monkey kidney cells were prepared and covered with an agarose overlay. Wells were formed in the agarose, the well bottom being optimally determined to be 3 mm from the monolayer and homotypic enterovirus antiserum, intersecting antiserum pool or antiserum‐diluent as control was added. After 2 h at 37°C, the test virus isolate was added to each well. Cultures were incubated at 37°C and were examined daily until cytopathic effect was readily observed in the control well. Monolayers were fixed and stained for macroscopic reading. Enterovirus identity based on the inhibition of cytopathic effect was confirmed with a conventional micro‐neutralization method. In all, 51 enterovirus isolates were typed. Included were 21 polioviruses, 9 coxsackieviruses and 21 echoviruses, all of which were correctly identified. This method takes advantage of the ability of enterovirus and antibody to diffuse through agarose. It is simple to perform. It does not require preliminary titration of the test virus isolate and tolerates 1,000 fold fluctuations in virus concentration, thereby offering laboratories a more rapid and efficient means of typing enteroviruses.
Lack of evidence of occult human immunodeficiency virus in seronegative individuals at very high risk of infection
Authors: Crowe Suzanne M. , Elbeik Tarek , Ulrich Paul P. , Mills John , Moss Andrew
Published: March 2007

Reports of human immunodeficiency virus (HIV) infection in seronegative individuals prompted the authors to investigate subjects who are at very high risk of acquisition of HIV in San Francisco. Nine HIV seronegative subjects were evaluated extensively, eight of whom were drawn from a well‐characterized cohort of seropositive and seronegative homosexual men who have been followed prospectively since 1983‐1984. These men have calculated probabilities of infection based on a fitted model of between 0.22 and 0.94. One additional subject is an intravenous drug user who has shared needles with HIV‐infected individuals extensively. Peripheral blood lymphocytes and monocytes were separately cultured from each subject and evidence of HIV infection was sought by a reverse transcriptase assay, enzyme immunoassay, and immunocytofluorographic analysis for HIV antigens, in situ hybridization, RNA slot blot analysis, and polymerase chain reaction amplification of HIV cDNA. Uncultured monocytes and lymphocytes from each donor were also examined by these techniques. Evidence of HIV infection was not found in the peripheral blood mononuclear cells of these high risk individuals.
Effects of purine nucleoside analogues with a cyclobutane ring and erythromycin A oxime derivatives on duck hepatitis B virus replication in vivo and in cell culture and HIV‐1 in cell culture
Authors: Hung Li‐Fang , Brumbaugh Amy E. , Bhatia Gulshan , Marion Patricia L. , Hung Paul P. , Norbeck Daniel W. , Plattner Jacob J. , Robinson William S.
Published: March 2007

The effects on duck hepatitis B virus (DHBV) replication of specific analogues of two classes of chemical compounds not previously tested against hepadnaviruses are described. One is erythromycin A‐9‐methyloxime (EMO) and other oxime derivatives of erythromycin A, and the other is purine nucleoside analogues (cyclobut A and cyclobut G)with cyclobutane rings. Viral replication was assessed by measuring serum levels of DHBV DNA in infected ducklings and DHBV DNA in infected primary duck hepatocyte cultures.

Administration of EM0 15 mg/kg of body weight IM to infected ducklings resulted in a rapid fall in DHBV DNA levels during therapy and a return to pretreatment levels after EM0 administration was stopped. There was local toxicity at injection sites with muscle necrosis in some animals. When 100 mg/kg EM0 was administered by gastric tube no such viral response was observed. The difference in virus response to EM0 15mgikg IM and 100 mgikg by gastric tube was not due to failure to achieve comparable blood and tissue levels of EM0 administered by the different routes. The results suggest an indirect effect dependent on IM injection of EM0 rather than a direct antiviral effect of the compound.

Administration of cyclobut G or cyclobut A at 70 mg/kg IM led to a rapid reduction of DHBV DNA to undetectable levels in serum, and in only 1 of 4 animals did DHBV DNA became detectable again within 10 days after stopping the drug. lntragastric administration of cyclobut G at 180 mg/kg per day was also associated with rapid reduction of serum DHBV DNA with onset of treatment in all animals, undetectable levels in 4 of 5 animals during therapy, and return to pretreatment levels in all animals after the drug was stopped. There were no apparent toxic effects. In cell culture 1 μM cyclobut A or cyclobut G inhibited virus production by more than twentyfold without apparent toxic effects. The results indicate that cyclobut A and cyclobut G are more active on a molar basis than other compounds tested against DHBV in these models, and they appear to be effective byu the oral route. Cyclobut G was much less active than AZT in inhibiting HIV‐1 in cell culture.
Survival of HIV‐1 activity after disinfection, temperature and pH changes, or drying
Authors: Tjøtta Enok , Hungnes Olav , Grinde Bjørn
Published: March 2007

A recently developed assay for measuring infectious HIV‐1 particles was used to determine the stability of the virus under various storage conditions as well as the effect of commonly used disinfectants. At the optimum pH of 7.1 the half life of the virus ranged from approx. twenty‐four hours at 37°C to no significant loss over 6 months at −75°C. Drying the virus on a glass surface or freezing caused a 5‐12 fold and 4‐5 fold decrease of activity, respectively. The dried preparations, however, were about as stable as when stored in a buffered solution. A solution of iodine and detergent (2% Jodopax) was the only disinfectant examined which removed all detectable HIV‐1 activity. Isopropanol and ethanol were more potent than acetone; however, all three solvents left some viable particles after a 30 min treatment with 70% solutions.
Differential antibody responsiveness to p19 Gag results in serological discrimination between human T lymphotropic virus type I and type II
Authors: Lal Renu B. , Brodine Stephanie K. , Coligan John E. , Roberts Chester R.
Published: March 2007

A new algorithm based upon the differential antibody responses to two gag gene products (p19 and p24) of human T lymphotropic virus (HTLV) has been suggested for serologic discrimination of HTLV type I (HTLV‐I) and type II (HTLV‐II) [Lillihoj et al., 1990]. To evaluate the practical usefulness of this algorithm, serum specimens from HTLV‐seropositive individuals whose infection was confirmed by PCR analysis to be HTLV‐I (n = 60) or HTLV‐II (n = 61) were analyzed by western blot. The intensities of the antibody response to p24gagand p19gag were scored by one individual without prior knowledge of PCR results. According to the algorithm, specimens with p19 > p24 were classified as HTLV‐I, whereas specimens with p19 < p24 were classified as HTLV‐II. Of 60 PCR confirmed HTLV‐I specimens, 56 had p19 > p24 (93%) while 4 had p19 < p24. Of 61 PCR confirmed HTLV‐II specimens, 56 had p19< p24 (92%) and 5 had p19 > p24. The overall accuracy of serologic differentiation when using this algorithm was 92%, as 4 of 60 HTLV‐I (7%) and 5 of 61 HTLV‐II (8%) could have been wrongly classified. Although the differential antibody response to p19gag and p2 4gag provides a simple means of serologically distinguishing between HTLV‐I and HTLV‐II infection in population‐based epidemiological studies, in a clinical context more accurate means of confirmation are required. The dominant p19gag responses were mapped to the C‐terminus of p19 (p19102–117) Competitive inhibition of p19gag response by peptide p19102–107, however, did not abrogate the binding of serum specimens from HTLV‐I‐infected individuals.
Low prevalence of hepatitis C virus and infrequent perinatal or spouse infections in pregnant women in Taiwan
Authors: Lin Ho‐Hsiung , Hsu Hong‐Yuan , Chang Mei‐Hwei , Hong Kun‐Fun , Young Yin‐Chin , Lee Tzu‐Yao , Chen Pei‐Jer , Chen Ding‐Shinn
Published: March 2007

To assess the prevalence of hepatitis C virus (HCV) infection in the pregnant women in Taiwan, we investigated two groups of pregnant women, 944 women without serum alanine aminotransferase (ALT) screening (group A) and 197 women with abnormal ALT (>45 IU/L) (group B). They were checked for anti‐HCV (anti‐C100‐3) with HCV EIA kit (Abbott Lab., North Chicago, IL). The results showed that 21 (2.2%) in group A and 5 (2.5%) in group B were anti‐HCV‐positive. However, 15 out of 21 in group A had an optical density (O.D.) of anti‐HCV <1.0, were negative by recombinant immunoblot assay (RIBA), and were regarded as false‐positive.

Nine infants delivered by those 11 cases were negative for anti‐HCV at 6 months of age, while none of the 8 husbands were anti‐HCV‐positive. It is concluded that the prevalence of anti‐HCV in pregnant women in Taiwan is low (6/944,0.63%), even in the cases with abnormal ALT (5/197, 2.5%). There was no serologic evidence for perinatal transmission or spouse infection.
Respiratory tract infections due to different rhinovirus serotypes and the influence of maternal antibodies on the clinical expression of the disease in infants
Authors: Kellner Gabriele , Popow‐Kraupp Therese , Binder Christa , Goedl Irene , Kundi Michael , Kunz Christian
Published: March 2007

Rhinoviruses were isolated from nasopharyngeal secretions of 49 children hospitalized because of severe respiratory tract infection. The isolates were typed using 90 type‐specific antisera. No obvious relation between certain serotypes and the severity of illness was found.

Serum samples were drawn from all children simultaneously with the nasopharyngeal secretions and screened for the presence of type‐specific neutralizing antibodies. Children aged 1 week t o 6 months had higher neutralizing antibody titers and revealed a lower degree of morbidity than older children. The decline of neutralizing serum antibodies with increasing age was correlated with a higher incidence of severe disease in those aged 7‐12 months. Nevertheless, also in this age group children with mild clinical courses of disease were observed despite a low concentration or an absence of neutralizing serum antibodies. This indicates that not only neutralizing serum antibodies, but other factors also influence the clinical expression of RHV‐induced disease.
Comparative evaluation of the immunogenicity of yeast‐derived (recombinant) and plasma‐derived hepatitis B vaccine in infants
Authors: Panda Subrat K. , Ramesh Rajagopal , Rao Kanury V. S. , Gupta Asha , Zuckerman Arie J. , Nayak Nabeen C.
Published: March 2007

The immunogenicity of plasma‐derived (HB Vax, MSD) and recombinant hepatitis B virus (Engerix B, SKF) vaccines was evaluated in infants born to hepatitis B virus carrier mothers. The vaccination was carried out at 1 day, 1 month, and 6 months of age using 10 μg of the vaccine given intramuscularly. A total of 83/88 (94.3%) and 74/79 (93.6%) of the infants receiving the plasma‐derived vaccine and yeast‐derived vaccine showed antibody to hepatitis B surface antigen (anti‐HBs). None of the maternal factors studied apart from the HBeAg positivity corellated with vaccine failure. The yeast‐derived vaccine gives marginally lower antibody titre than the plasma‐derived vaccine. The group‐specific anti‐“a” antibody was less than 10% of the total anti‐HBsAg titre. It was observed that the vaccine alone without prior administration of hepatitis B immunoglobulin is effective in perinatal infection.
Human T‐cell lymphotropic virus type 1 infection and tropical spastic paraparesis in belgian expatriates
Authors: Goubau P. , Carton H. , Cornet P. , Vercauteren G. , van Gompel A. , de Vooght H. , Piot P. , Desmyter J.
Published: March 2007

A case of HTLV‐1 associated tropical spastic paraparesis is described in a Belgian nun who had been working as a midwife in Central Africa. Occupational exposure was the only risk factor identified. Among 2,482 Belgian expatriates in tropical countries, 92% of whom had resided in sub‐Saharan Africa for an average of 15.5 years, only one Belgian‐born man was found seropositive for HTLV‐1. He was married t o an African woman and living in Central Africa for 23 years. The risk of HTLV‐1 infection is low in Belgian expatriates and on its own does not support generalised anti‐HTLV screening in autochthonous Belgian blood donors.
Transfusion‐acquired cytomegalovirus infection in children in a hyperendemic area
Authors: Lee Ping‐Ing , Chang Mei‐Hwei , Hwu Wuh‐Liang , Kao Chuan‐Liang , Lee Chin‐Yun
Published: March 2007

Thirty‐nine children without previous cytomegalovirus (CMV) infection received blood transfusion in the National Taiwan University Hospital. The overall transfusion‐acquired CMV infection rate was 36% (14/39). Donor CMV seropositive rate was 70%. None of the nine children who had received seronegative blood became infected, in contrast to 14 of the 21 children (67%) who had received seropositive blood (P = 0.002). Another significant risk factor associated with CMV infection was the use of fresh blood: 13 of 15 (87%) with fresh seropositive blood were infected, in contrast to one of six (17%) with “old” seropositive blood (P = 0.01). Most of the fresh blood was used within 24 hours. This blood processing method was shown to account for the extremely high rate of CMV infection in those who had received fresh seropositive blood. The results indicated that the incidence of CMV infection can be reduced by avoiding the use of fresh blood, especially blood less than 24 hours old. For such a population in Taiwan with high prevalence of positive CMV antibody, this approach was more applicable than screening donor blood for CMV antibody.
Antibodies to Epstein‐Barr virus and cytomegalovirus in relation to CD4 cell number in human immunodeficiency virus 1 infection
Authors: Quesnel A. , Pozzetto B. , Touraine F. , Moja P. , Lucht F. , de The G. , Touraine J. L. , Gaudin O. , Genin C.
Published: March 2007

Interaction between herpesviruses and human immunodeficiency virus (HIV)I is postulated in the progression of HIV disease. In order to evaluate the specific antibody responses directed to Epstein‐Barr virus (EBV) and cytomegalovirus (CMV) and to provide serological evidence suggesting reactivation of these viruses able to accelerate the immunodeficiency, we studied IgA and IgG titres to EBV and CMV in the serum of HIV positive patients in relation to the CD4 cell number.

The titres of IgG antibodies to EBV and the prevalence of IgG to CMV were significantly higher i n HIV positive patients compared to control high risk HIV negative subjects. In HIV infected patients, anti‐VCA IgG antibodies increased and anti‐EBNA IgG antibodies decreased progressively in relation t o the decline of D4 cell number whereas anti‐CMV IgG antibodies did not varied significantly at the same time. Anti‐VCA IgA and anti‐EA IgG antibodies were found uncommonly and with low titres. IgA antibodies to EA and CMV were not detected in any patient. The variations in EBV antibody response that we describe in HIV infection were previously reported in other immunodeficiency states and could be distinctive of these diseases.
Replication of hepatitis B virus in differentiated adult rat hepatocytes transfected with cloned viral DNA
Authors: Diot Christian , Gripon Philippe , Rissel Maryvonne , Guguen‐Guillouzo Christiane
Published: March 2007

The possibility of obtaining expression of human hepatitis B virus (HBV) genes and production of virus particles in normal liver cells from heterologous species like normal adult rat hepatocytes, by transfecting the complete HBV genome, was investigated. Various techniques for hepatocyte transfection were assayed including the usual calcium‐phosphate coprecipitation technique, the Pasco and Fagan modified calcium‐phosphate procedure, and the lipofection technique. Transfection efficiency was determined by measuring the production of HBV surface antigen under various culture conditions. Transfection was the most efficient when assayed 1 or 2 days after hepatocyte plating at low density. Few variations in the efficiency were observed between the different transfection procedures. We show that under these culture conditions, replication of HBV can be achieved in differentiated adult rat hepatocytes. Synthesis of relaxed circular and single‐stranded DNA forms and of viral transcripts including pregenome RNA occurred in the cells whereas viral antigens and mature and immature viral particles were released into the culture medium. The production of viral proteins was always higher in hepatocytes cocultivated with rat liver epithelial cells and maintained at a low density. In contrast, viral replication was not obtained by transfecting undifferentiated rat liver epithelial cells. These results demonstrate that replication of HBV can occur in hepatocytes from mammalian species non‐closely related to primates and strongly support the idea that attachment of the virus and its penetration into the cells are critical steps in the host‐specificity of the infection process and that hepatic‐specific regulating factors could be essential for viral replication. This model provides a powerful experimental system which is easy to use t o test the role of HBV DNA sequences on normal hepatocyte functional activity, particularly those expected to transactivate cellular genes.
Increased detection of HPV 16 virus in invasive, but not in early cervical cancers
Authors: Labeit D. , Back W. , Weizsäcker F. V. , Hermann P. , Melchert F. , Leonard K. , Labeit S.
Published: March 2007

Human papilloma virus type 16 (HPV 16) DNA is found in about 50% of cervical squamous cell carcinomas (SCCs), and this association has raised the possibility of a causal role for HPV 16 in cervical carcinogenesis. We have tested this hypothesis by assaying a series of biopsies (n = 119) ranging from normal mucosa to infiltrating SCC with the PCR‐technique for the presence of HPV 16 DNA. While HPV 16 DNA was detected in 50% of our cases with invasive SCC, the incidence of HPV 16‐positive samples was about 10% in all other biopsies ranging from normal mucosa t o cases of carcinoma in situ. HPV 16 therefore appears to be involved in late tumor promotion but not in early tumor development.
Lack of detectable human T‐cell lymphotropic virus type I sequences in samples from multiple sclerosis patients
Authors: Menzo Stefano , Manzin Aldo , Bagnarelli Patrizia , Varaldo Pietro E. , Grandi Giorgio , Giuliani Giorgio , Cazzato Giuseppe , Giacca Mauro , Clementi Massimo
Published: March 2007

Recently, inconclusive results have followed the early data on the possible association between multiple sclerosis (MS) and human T‐cell lymphotropic virus type I (HTLV‐I) infection. For this reason, we examined this hypothesis using the polymerase chain reaction (PCR) to study samples of differing origin from Italian MS patients.

In particular, we developed a systematic analysis of paraffin‐embedded brain white matter from histologically defined lesions of 14 MS patients using PCR and primer sets specific for HTLV‐I sequences; additionally, cerebrospinal fluids (CSFs) from 12 patients and peripheral blood mononuclear cells (PBMCs) from subjects at the early and late phase of the disease were investigated for free HTLV‐I virions and specific proviral sequences, respectively.

In agreement with some groups who reported lack of HTLV‐I sequences in PBMCs of MS patients but in clear contrast with others, we failed to detect specific viral sequences using this broad approach.
Application of screening and confirmatory assays for anti‐HTLV‐I/II in U.K. populations
Authors: Tosswill Jennifer H. C. , Parry John V. , Weber Jonathan N.
Published: March 2007

During an epidemiological study of a low risk U.K. population diverse screening and confirmatory assays for the detection of anti‐HTLV‐1/11 were assessed. Sera from 2,900 antenatal patients were tested for anti‐HTLV‐1/11 by gelatin particle agglutination assay (GPA). All reactive sera, and 133 randomly selected unreactive sera, were further tested by Abbott and DuPont ELISAs, “in house” competitive and IgG capture radioimmunoassays (RIAs), and Western blot (WB). Sera which reacted with any HTLV‐I proteins by WB were tested by radio immunoprecipitation assays (RIPA). The two ELlSAs detected all the GPA reactive specimens that were subsequently confirmed as anti‐HTLV‐1/11 positive. Confirmation of positive screening results required the use of both WB and RIPA. Serological diagnosis of HTLV infection involves access to specialised assays that are not commercially available.
Salivary antibody titers in adults challenged with a human rotavirus
Authors: Ward Richard L. , Pax Kristine A. , Sherwood James R. , Young Elizabeth C. , Schiff Gilbert M. , Bernstein David I.
Published: March 2007

To determine whether salivary antibody is a reliable indicator of rotavirus infection and mucosal rotavirus antibody concentrations, salivary rotavirus antibody titers were determined as a function of time following inoculation of 24 adult volunteers with a virulent strain of human rotavirus (CJN). Twenty of the subjects became infected and all produced detectable amounts of salivary rotavirus IgA. These antibody concentrations remained undetectable in the four uninfected subjects. Rises of ≥ fourfold in either salivary rotavirus IgA or neutralizing antibody to the challenge virus were detected in all but two subjects. The titers of both antibodies were maximal at 13 days after inoculation and decreased significantly (p < .02) by 27 days, a result very similar to that previously found with stool rotavirus IgA in CJN‐infected subjects. These results suggest that salivary rotavirus antibody titers are an accurate reflection of mucosal rotavirus anti‐body and a possible surrogate for intestinal rotavirus antibody.
Temporal relationships of hepatitis C virus RNA and antibody responses following experimental infection of chimpanzees
Authors: Beach Michael J. , Meeks Emory L. , Mimms Larry T. , Vallari David , Ducharme Louise , Spelbring John , Taskar Suhas , Schleicher Joseph B. , Krawczynski Krzysztof , Bradley Daniel W.
Published: March 2007

Liver enzyme levels, viral RNA, and the immune response against both structural and nonstructural hepatitis C virus (HCV) proteins have been studied in experimentally infected chimpanzees in order to further understand the natural history of HCV infection. An ELISA for measuring both IgG and IgM responses to core (c22), 33c (NS3), and c l00. (NS4) was employed. The IgG response rates were 5/8 for core, and 8/8 for both 33c and c100. Utilizing this antigen combination, a t least one antibody response is measurable at, or within 3 weeks of, the major ALT peak. Although no individual antibody response is universally associated with initial detection of seroconversion, the combination of all three recombinant proteins measures seroconversion an average of 54 days earlier than with c100 alone, in 6/8 of the animals. IgM responses were measurable in 5/8 of the chimpanzees, were of shorter duration, and usually arose concomitantly with IgG responses. IgM appears to be a good indicator of primary infection since neither boosting nor recrudescence of disease during the chronic phase of disease elicited a secondary IgM response. Viral RNA can be measured 4‐7 days (average = 9 days) postinfection with the period preceding the ALT peak being characterized by several PCR positive segments interrupted by periods in which no viral RNA can be measured. Following the ALT peak, chronically infected animals with recurring ALT elevations are generally PCR positive with intercedent PCR negative periods. Those animals that appear to have biochemically resolved disease generally have PCR negative profiles, although they still may periodically exhibit PCR positive sera. This indicates that with the recent advent of new screening techniques, a more stringent definition of HCV resolution will be required.
Association of hepatitis C virus carrier state with the occurrence of hepatitis C virus core antibodies
Authors: Claeys Hendrik , Volckaerts André , Beenhouwer Hans D. , Vermylen Carl
Published: March 2007

An enzyme immunoassay (EIA) was developed for the determination of antibodies against the “putative” core protein of hepatitis C virus (HCV). Antigens used were recombinant fragments (amino acids 6‐77 or 6‐143) of the HCV core protein, produced in Escherichia coli with truncated hepatitis B core (HBc) as fusion protein.

Evaluation of 385 sera positive for HCV antibodies by first generation EIA, revealed 98 (25.4%) with HCV core antibodies. HCV‐RNA, determined by the polymerase chain reaction (PCR), was exclusively found in the sera positive for HCV core antibodies (89 PCR positives). In random screening of 3,708 sera, 3 sera with HCV core antibodies were found PCR positive. Only 2 of these sera were positive in the first generation EIA

It is concluded that HCV core antibody determination is a reliable test for identifying HCV carriers among blood donors.
Prevalence of high risk genital papillomaviruses in the Belgian female population determined by fast multiplex polymerase chain reaction
Authors: Vandenvelde C. , Scheen R. , van Pachterbeke C. , Loriaux C. , Decelle J. , Hubert T. , Delhaye C. , Cattoor J.‐P. , Buys M. , van Beers D.
Published: March 2007

Because in situ/filter hybridisation is not sensitive enough and because classical polymerase chain reaction (PCR) protocols are generally not sufficiently reproducible and specific, there is little accurate information on the prevalence of human papillomaviruses (HPV) 16,18, and 33 infections in women without dyskaryotic changes of the cervix.

In our hands, our Fast Multiplex PCR protocol has always been the most sensitive, specific, and reproducible DNA detection assay in all the microbiological and haematological applications we attempted (Vandenvelde C, Verstraete M, Van Beers D [ 1990] : Journal of Virological Methods 30:215‐228; Vandenvelde C, Scheen R, Corazza F, Van Beers D [1991a]: Journal of Experimental and Clinical Hematology 33:293‐297;Vandenvelde C, Scheen R, Van Beers D, Fondu P [1991 b]: Journa lof Experimental and Clinical He‐matology 30:25‐29). Using this new technique, cervical scrapes from 336 Belgian women attending the cervical cancer screening clinic were examined for the presence of these three high‐risk genital papillomaviruses. Positive results were confirmed using another set of HPV‐specific primers. Exactly one sixth of our population was found positive for one or more of these HPVs. Types 33 and 16 were significantly more prevalent than type 18. The nonparametric statistical analysis of the data suggests that some risk factors such as particular sexual habits, that are inversely related to age, must exist.
Prevalence of human papillomavirus infections among heterosexual men and women with multiple sexual partners
Authors: van Doornum Gerard J. J. , Hooykaas Christa , Juffermans Leon H. J. , van der Lans Sylvia M. G. A. , van der Linden Mireille M. D. , Coutinho Roel A. , Quint Wim G. V.
Published: March 2007

A prospective study of 65 men and 111 women with multiple heterosexual partners was designed to assess the prevalence and potential risk factors of genital human papillomavirus (HPV) infections. In addition, the HPV reservoir in genital, rectal, and oral mucosa was examined.

The specimens for the detection of HPV DNA were taken from different sites such as the urethra and coronal sulcus (men), cervix and labia minora (women), anus, rectum, tongue, and buccal mucosa (both men and women). Women underwent speculum examination and colposcopic evaluation of the anogenital region, and a smear for routine cytological classification was also taken. In men, the anogenital region was examined clinically and colposcopically.

The polymerase chain reaction (PCR) was used for the detection of HPV types 6/11, 16, 18, and 33.

A high prevalence of HPV infection at one or more sites was detected, in 32% of the male and in 23% of the female participants. Seventeen percent of the male distal urethral specimens were positive for HPV DNA. From the female cervical specimens 14% were found positive. Ten proctal specimens (five men and five women) were positive for HPV DNA without any discernible lesion. The persons from whom these samples were taken denied anal insertive intercourse. No oral manifestation of HPV infection was detected.

In both men and women a difference between HPV DNA‐positive and ‐negative persons was not found in relation to known risk factors associated HPV infection.

The clinical manifestation of HPV infections in this group of heterosexual men and women with multiple sexual partners is characterised by a high rate of latency both in men (72%) and women (80%), which may be important in the understanding of the transmission of HPV. The male urethra was identified as an important site of latent infection. The data support heterosexual transmission of HPV and suggest a nonsexual means of spread at the same time. © 1992 Wiley‐Liss, Inc.
Seroepidemiology of cytomegalovirus infection among children between the ages of 4 and 12 years in Taiwan
Authors: Shen Chen‐Yang , Chang Whi‐Wen , Chang Shu‐Fen , Chao Maio‐Fen , Huang Eng‐Shang , Wu Chen‐Wen
Published: March 2007

To determine risk factors responsible for primary cytomegalovirus (CMV) infection in Taiwan, samples of blood for antibody to CMV were obtained from 362 children aged 4 to 12 years: 58% were found to be positive for anti‐CMV IgG antibody. Logistic regression analysis showed that seropositivity correlated with age, method of delivery, duration of breast feeding, and younger age of mother. Neither socioeconomic status nor crowded living conditions showed significant correlation with CMV seropositivity. Primary CMV infection in Taiwan appears to be less related to socioeconomic status shown in western countries, and, since the majority of pregnant women were seropositive in Taiwan, two of the major sources of primary CMV transmission are infected breast milk and the infected genital tract. © 1992 Wiley‐Liss, Inc.
Detection of the human papilloma virus type 16 mRNA‐transcripts in cytological abnormal scrapings
Authors: Falcinelli Cristina , Claas Eric , Kleter Bernhard , Quint Wim G. V.
Published: March 2007

The RNA‐polymerase chain reaction (PCR) was carried out on cervical scrapings to detect and analyze transcripts from the E6‐E7 open reading frames (ORF) of human papilloma virus type 16 (HPV16). The method, described previously for cervical squamous carcinomas and cervical intraepithelial neoplasias, was adapted to cervical scrapings. A primer set and two different probes specific for the E6‐E7 ORFs were selected. One of the probes was able to detect the amplification products from the full length, the major, and the minor transcripts whereas the other was specific for the major transcript only. To check the quality of the mRNA in the cervical scrapings, a primer set and a probe specific for the human keratin mRNA were selected. A group of 17 abnormal cytological cervical scrapes, which were positive for HPV16 DNA, was analyzed.

In this group the human papilloma virus was not always transcriptionally active, as HPV16 mRNA transcripts were detected only in about one‐half (8/17) of the samples. These findings suggest that the RNA‐PCR method on cervical scrapings may be very useful for epidemiological studies on the role of transcriptionally active/ inactive HPV16 genes in the pathogenesis of an HPV16 infected lesion. © 1992 Wiley‐Liss, Inc.
Low antibody avidity in elderly chickenpox patients
Authors: Schoub Barry D. , Blackburn Nigel K. , Johnson Sylvia , McAnerney Jo M. , Miller Bennie
Published: March 2007

A small outbreak of chickenpox confirmed serologically in 3 elderly patients from a geriatric home is described. Disease was probably due to exogenous reinfection, yet nevertheless the avidity of specific antibodies measured by the urea denaturation test was even lower than in primary chickenpox controls, which themselves were, as expected, significantly lower than zoster controls. In elderly individuals susceptibility to reinfection with varicella‐zoster virus (VZV) with clinical manifestation such as chickenpox may well be associated with the decay of specific humoral immunity detectable by antibodies of particularly low avidity, in contrast to reactivation of latent VZV presenting clinically as zoster, which is related to deficiencies in specific cellular immunity. © 1992 Wiley‐Liss, Inc.
Investigation of an outbreak of rotavirus infection in geriatric patients by serotyping and polyacrylamide gel electrophoresis (PAGE)
Authors: Steel H. M. , Garnham S. , Beards G. M. , Brown D. W. G.
Published: March 2007

An outbreak of rotavirus diarrhoea amongst pa‐tients on two wards in a geriatric unit was inves‐ tigated using polyacrylamide gel electrophoresis (PAGE) and serotyping. There had been no con‐ tact between the patients on different wards but some interchange of staff had occurred so it was important to exclude the possibility that the rota‐virus had been spread by staff.

Two strains of rotavirus were shown to be present, one on each ward. Differences in the electropherotypes and serotypes were demon‐strated, showing conclusively that transfer of vi‐rus between the wards had not occurred. Thus, serotyping and PAGE are of value in the investi‐gation of rotavirus outbreaks in the hospital setting where cross‐infection between wards is suspected. © 1992 Wiley‐Liss, Inc.
Detection of adenoviruses in stools from healthy persons and patients with diarrhea by two‐step polymerase chain reaction
Authors: Allard Annika , Albinsson Bo , Wadell Göran
Published: March 2007

The use of the polymerase chain reaction (PCR) for detection of human adenoviruses in diluted stool samples was investigated. Two sets of nested primers, including primers specific for the hexon‐coding region and for the E1 B region of enteric adenoviruses (EAd), were assessed by two‐step amplification. The primers constitute two different PCR systems designed for the detection of adenoviruses belonging to all six subgenera (A‐F), and the two EAds Ad40 and Ad41, respectively. In a two‐step PCR mediated amplification a single virus particle was detected when the two sets of general hexon primers or Ead specific primers were used. Earlier results from PCR detection of adenoviruses in stool from children suffering from diarrhea gave indications that adenovirus particles are commonly shed in stools without being identified as the cause of il I ness [Allard et a I.: Journal of Clinical Microbiology 28:2659‐2667, 1990]). Therefore, the general and the EAd specific PCR assays were assessed on 150 stool specimens from three groups including 50 healthy children, 50 healthy adults, and 50 adults suffering from diarrhea. When the two sets of general hexon primers were used, 25 of the 50 specimens from the healthy children (mean age 21 months) were found positive by two‐step PCR amplification. Nine of the 50 specimens from the healthy adults (mean age 32 years) were found positive whereas 12 of the 50 specimens from sick adults (mean age 31 years) gave amplification products, using the two sets of general hexon primers in a nested fashion. None of the 150 specimens were found to be positive by two‐step PCR amplification using the two sets of EAd‐specific primers. The sensitivity of the PCR, together with its simplicity and reduced time scale compared to other detection methods, support the potential of this technique as an additional method for routine detection of human adenovirus infections. © 1992 Wiley‐Liss, Inc.
Typing of human group A rotavirus with alkaline phosphatase‐labeled oligonucleotide probes
Authors: Sethabutr Orntipa , Hanchalay Suvath , Lexomboon Udom , Bishop Ruth F. , Holmes Ian H. , Echeverria Peter
Published: March 2007

Rotavirus (RV) in stools of children <1 year of age with diarrhea in Bangkok in 1989 were serotyped by monoclonal enzyme immunoassay (MEIA). RNA extracted from these specimens was tested for hybridization with alkaline phosphatase (AP) and 32P‐labeled oligonucleotides constructed from the nucleotide sequences of VP7 of human G types 1 (HuG1Ac), 2 (HuG2Ac), 3 (HuG3Ac), and 4 (HuG4Ac). Of 148 specimens that contained RV, 72% (106/148) hybridized with RV G type specific AP‐labeled oligonucleotides compared to 47% (70/148) that were serotyped by MEIA (P < 0.001). Of 68 specimens that contained only one VP7 serotype (G‐type), as identified by MEIA, 94% (16/17) of G1, 90% (27130) of G2, 57% (4/7) of G3, and 36% (5/14) of G4 RV hybridized with the AP‐labeled HuG1Ac, HuG2Ac, HuG3Ac, and HuG4Ac oligonucleotides, respectively. The probes for G1, 2,3, and 4 RV were specific for each G type. The results of hybridizing specimens with 32P‐ and AP‐labeled oligonucleotides were similar. After transcription and amplification of cDNA of gene 9, AP‐labeled RV G type specific oligonucleotides hybridized with 90% (134/148) of RV specimens. The high sensitivity of these nonimmunological techniques could be of value in identifying G types of RV during vaccine trials. © 1992 Wiley‐Liss, Inc.
A microtitre format point mutation assay: Application to the detection of drug resistance in human immunodeficiency virus type‐1 infected patients treated with zidovudine
Authors: Kaye Steve , Loveday Clive , Tedder Richard S.
Published: March 2007

An assay for the analysis of point mutations in DNA fragments amplified by the polymerase chain reaction is described. The method was applied to the analysis of mutations leading to single amino acid changes in the reverse transcriptase gene of human immunodeficiency virus type‐1 that are associated with decreased sensitivity of the virus to the effects of the nucleoside analogue zidovudine. The assay uses a mi‐crotitre format allowing large numbers of clinical samples to be analysed, and is a quantitative assay giving information on the relative proportions of wild‐type and mutant sequence at the point being analysed. The analysis of mutations in codons 67, 70, 215, and 219 of the RT gene in samples taken at various time points after the commencement of zidovudine therapy shows a high variability in the rate at which these mutations arise and in the proportions of wild‐type and mutant sequences observed. The method we describe has wide application to the analysis of other point mutations in clinical or research Studies. © 1992 Wiley‐Liss, Inc.
Hepatitis B surface antigen particles of subtypes adw and adr, and compound subtype (adwr) in symptom‐free carriers in Japan
Authors: Kanagawa Hiroshi , Takai Emiko , Tsuda Fumio , Machida Atsuhiko , Kojima Mineo , Ishijima Ayako , Tanaka Takeshi , Okamoto Hiroaki , Miyakawa Yuzo , Mayumi Makoto
Published: March 2007

Of sera from 1,878 Japanese blood donors who carried hepatitis B surface antigen (HBsAg), 420 were subtyped as adw(22.4%) and 1,443 as adr(76.8%); only 15 (0.8%) contained HBsAg of subtype aywor ayr. Sera with HBsAg/adrhad higher HBsAg titres than those with HBsAg/adw (geometric mean of haemagglutination titre: 10.1 ± 2.4 vs. 9.7 ± 2.4, p <0.01), and a higher prevalence of hepatitis B e antigen (24% vs. 13%, p <0.001). Carriers of HBsAg/awr progressively predominated over those of HBsAg/adwwith increasing age. Of sera from 1,863 carriers of HBsAg/adwor HBsAg/adr, 182 (9.8%) contained HBsAg particles with both subtypic determinants in the wir allele. The presence of wand rdeterminants on the same particles was ascertained by sandwiching them between monoclonal antibody with the specificity for w and that with the specificity for r. HBsAg particles of compound subtype (adwr)were found more often in sera with hepatitis B e antigen than those without it (145/403 [36.0%] vs. 37/1,460 [2.5%], p <0.001). Sera with HBsAg/adwrparticles had HBsAg titres higher than those without them (12.4 ± 1.9 vs. 9.7 ± 2.3, p <0.001). HBsAg/adwrparticles arise from phenotypic mixing of the S‐gene product of wild‐type virus and that of mutants with point mutations for subtypic changes. The results obtained indicated that HBV strains of subtype adrhave a higher replicative activity than those of adw, and suggested that mutations in the S gene for subtypic changes would be associated with an active replication of hepatitis B virus. © 1992 Wiley‐Liss, Inc.
Concurrent hepatitis C virus and hepatitis delta virus superinfection in patients with chronic hepatitis B virus infection
Authors: Liaw Yun‐Fan , Chien Rong‐Nan , Chen Tong‐Jong , Sheen I‐Shyan , Chu Chia‐Ming
Published: March 2007

Since hepatitis C virus (HCV) and hepatitis delta virus (HDV) are transmitted by the same routes as hepatitis B virus (HBV), simultaneous or concurrent HCV and HDV infection in patients with chronic HBV infection may occur. To test this hypothesis and to examine the clinicohistological and immunopathological presentations of such multiple hepatitis virus infections, acute and/or convalescent serum specimens from 86 patients with acute HDV superinfection were tested by enzyme immunoassay for antibodies to HCV. Of the 86 patients, 18 (20.9%) were associated with HCV infection. Although patients with early mortality cannot be evaluated by the HCV markers used in this study, the results showed that the clinical and histologic features were similar except that patients with HCV infection were older than those without HCV infection (P < 0.01). Immunopathological studies carried out within 2 months after the onset of acute HDV superinfection demonstrated that hepatitis B core antigen (HBcAg) was not detected in any patient and HDV antigen was detected in 18.2% of the patients with HCV infection whereas HBcAg and HDAg were found in 7% and 65.1%, respectively, of those without HCV coinfection (P < 0.02). It is concluded that concurrent HCV and HDV superinfections can and do occur in patients with chronic HBV infection. In these triple viral infections, HCV may even transiently suppress HDV and HBV. © 1992 Wiley‐Liss, Inc.
Detection of hepatitis “C” virus in formalin‐fixed liver tissue by nested polymerase chain reaction
Authors: Sallie Richard , Rayner Anne , Portmann Bernard , Eddleston A. L. W. F. , Williams Roger
Published: March 2007

Interpretation of antibody to hepatitis C virus (HCV) in patients with liver disease is difficult due to false‐positive reactivity in some conditions. To evaluate the feasibility of HCV in archival material, HCV was sought in formalin‐fixed, paraffin‐embedded liver biopsy specimens. Nested polymerase chain reaction was used to detect hepatitis C virus in formalin‐fixed, paraffin‐embedded liver biopsy specimens after total RNA was extracted from tissue by proteinase K digestion and phenol/chloroform purification. The relative efficiency of amplification of HCV RNA from formalin‐fixed material was estimated semiquantitatively by serial dilution of cDNA synthesised from RNA extracted from fresh and formalin‐fixed sections from the same liver. Although HCV RNA could be detected in formalin‐fixed liver tissue by nested PCR in 5/5 cases in which HCV was detected in serum, amplification was ∼5‐fold less efficient than when HCV was amplified from fresh tissue. Nevertheless, nested PCR of HCV from formalin‐fixed liver tissue represents a useful technique in addressing some important questions related to the pathogenesis of liver disease. © 1992 Wiley‐Liss, Inc.
Prevalence of serum antibodies against lymphocytic choriomeningitis virus in selected populations from two U.S. cities
Authors: Stephensen Charles B. , Blount Sharon R. , Lanford Robert E. , Holmes Kathryn V. , Montali Richard J. , Fleenor Michael E. , Shaw Joseph F.E.
Published: March 2007

An ELISA was developed for measuring serum antibodies against the arenavirus lymphocytic choriomeningitis virus (LCMV) and a closely related isolate termed callitrichid hepatitis virus (CHV). The ELISA was used to test sera from healthy adults and from hepatitis patients. In Birmingham, Alabama, the seropositivity rate for healthy black women was 5.1% (7/138), and the rate for patients with all types of hepatitis or cirrhosis was 4.3% (2/46). In San Antonio, Texas, the seropositivity rate among a clinical series of patients with non‐A, non‐B hepatitis was 0 (0/20), and the rate among persons rejected from blood donation because of high serum alanine aminotransferase levels was 2.4% (2/82). These results indicate that infection with LCMV or CHV is common in Birmingham but that infection is not associated with hepatitis. © 1992 Wiley‐Liss, Inc.