bluuebunny/crossref_metadata_2025 huggingface dataset:
Evaluation of a new generation synthetic peptide combination assay for detection of antibodies to HIV‐1, HIV‐2, HTLV‐I, and HTLV‐II simultaneously
Authors: Zhang Xiang , Constantine Niel T. , Bansal Jaya , Callahan Johnny D. , Marsiglia Vincent C.
Published: March 2007

A new generation combination test (Detect‐Plus, IAF BioChem, Montreal, Canada) based on synthetic peptides for HIV‐1, HIV‐2, HTLV‐I, and HTLV‐II was compared with three routine commercial screening assays and confirmatory assays to determine its sensitivity and specificity and to evaluate it as a substitute screening method. Samples from 356 sexually transmitted disease (STD) patients were tested by the four screening tests. All initially reactive samples were retested in duplicate by the corresponding EIA and repeatedly reactive samples were confirmed by Western blots for HIV‐1, HIV‐2, and HTLV‐1/11. The confirmed positives detected by each screening assay were HIV‐1 (23/356,6.46%), HIV‐2 (1 1/356,3.09%), and HTLV‐1/11 (5/356,1.4%). The new generation Detect‐Plus test produced only two results (2/356, 0.56%) that were presumed t o be false‐positives in comparison to the screening tests, but the OD/CO values were just slightly high (1.5 and 1.9). There were no false negative results, indicating that the sensitivity of the new combination test was excellent (100%). Compared with routine retroviral EIA assays, the test is easy to perform—the total time requirement is only 2 hr and there is no need for incubation equipment. The OD/CO values were very high when samples were positive, making even visual interpretation possible. We conclude that this new combination assay is an excellent screening method for detection of antibodies to the human retroviruses, and may be particularly useful for screening blood for transfusion and in epidemiological investigations. © 1992 Wiley‐Liss, Inc.
Effects of transforming growth factor‐β1 against the inhibitory action of interferon on DNA synthesis and viral replication in hepatitis B virus DNA‐transfected cell
Authors: Kakumu Shinichi , Ito Yuji , Wakita Takaji , Yoshioka Kentaro , Ishikawa Tetsuya , Takayanagi Masahiro , Higashi Yasuyuki , Yang Zhi‐Qi
Published: March 2007

Studies were undertaken to examine the effects of recombinant human transforming growth factor beta 1 (TGF‐PI) on DNA synthesis and antiviral actions of interferons (IFNs) in HepG2 cell, a hepatoma cell line, transfected with hepatitis B virus (HBV) DNA. The inhibitory effects of IFN‐a and ‐γ on DNA synthesis of HepG2 cells were enhanced in a dose‐dependent manner by a simultaneous addition of TGF‐β. The degree of suppression by the reagents was greater in HBV nontransfected cells than in transfected cells. Inhibition of DNA synthesis was not due to direct cytotoxic effects of the additives, since the viability of HepG2 cells was comparable i n the control and treated cultures as determined by trypan blue exclusion. Treatment of HBV DNA‐transfected HepG2 cells with IFNs resulted in decrease in production of HB surface and e antigens, and in the level of HBV DNA, but TGF‐p reversed the IFN‐induced antiviral state in HBV DNA‐transfected HepG2 cells. TGF‐p had no direct effect on HBV replication. These results indicate that rTGF‐p1 exerts a differential effect against the inhibitory actions of IFN on DNA synthesis and viral replication in HepG2 cells. © 1992 Wiley‐Liss, Inc.
Detection of hepatitis C viraemia in caucasian patients with hepatocellular carcinoma
Authors: Garson J. A. , Wicki A. N. , Ring C. J. A. , Joller H. , Zala G. , Schmid M. , Buehler H.
Published: March 2007

Potential risk factors for the development of hepatocellular carcinoma were analysed in 40 Caucasian patients with this malignancy. A higher proportion (14 of 40; 35%) had evidence of hepatitis C virus (HCV) infection than had evidence of either hepatitis B virus (HBV) carriage (17.5%) or alcohol abuse (30%). In all 14 patients whose sera were reactive by HCV ELISA (Ortho second generation test), the presence of antibodies to HCV were confirmed by recombinant immunoblot assay (Ortho RIBA‐2). Furthermore, two independent laboratories detected HCV‐RNA in 10 of the 14 (71%) anti‐HCV positive sera. Two additional sera were shown to contain HCV‐RNA when reanalysed by a modified PCR using oligo‐nucleotide primers designed to amplify a shorter fragment of the 5′ noncoding region of the genome. Seven of the anti‐HCV positive patients also had evidence of prior HBV infection and 2 admitted to alcohol abuse. HCV infection was the only identifiable risk factor in 6 patients. These data confirm the association between HCV infection and hepatocellular carcinoma and suggest that persistent viral replication accompanies tumour development in the majority of patients whose serum contains anti‐HCV. © 1992 Wiley‐Liss, Inc.
Dehydroepiandrosterone protects mice inoculated with West Nile virus and exposed to cold stress
Authors: Ben‐Nathan D. , Lustig S. , Kobiler D. , Danenberg H. D. , Lupu E. , Feuerstein G.
Published: March 2007

The protective effect of pretreatment with dehydroepiandrosterone (DHEA) on stress‐enhanced viral encephalitis was studied in mice exposed to cold following inoculation with West Nile virus (WNV). Exposure of WNV‐inoculated mice to cold water (1 ± 0.5°C, 5 minutes/day for 8 days) resulted in a mortality rate of 83% as compared to 50% in nonstressed mice (p < 0.05). The effect of cold stress was more pronounced when mice were inoculated with WN‐25, a noninvasive neurovirulent variant of WNV. Mice infected with WN‐25 showed no mortality, whereas cold stressed mice inoculated with the same virus had a mortality rate of 67% (p < 0.05). The administration of DHEA (serial injections of 10‐20 mg/kg with or without a loading dose of 1 gm/kg) resulted in a significant reduction in the mortality rate of stressed mice inoculated with either virus (p < 0.05). Virus levels in the blood and brain of the DHEA‐treated mice, were significantly lower than in the control groups.

DHEA also prevented the involution of lymphoid organs in stressed mice. The present study provides direct evidence of the protective effects of DHEA as an “anti‐stress” agent. Its ability to prevent mortality associated with WNV or WN‐25, and involution of lymphoid organs caused by stress‐induced immunosuppression, supports the notion that its activity is based on the modulation of the host response. © 1992 Wiley‐Liss, Inc.
Improved diagnosis of chronic hepatitis C virus infection by detection of antibody to multiple epitopes: Confirmation by antibody to synthetic oligopeptides
Authors: Brown D. , Powell L. , Morris A. , Rassam S. , Sherlock S. , McIntyre N. , Zuckerman A. J. , Dusheiko G. M.
Published: March 2007

Serum samples from 226 patients covering a wide spectrum of liver disease were tested for antibodies to hepatitis C virus (HCV) using both first and second generation enzyme linked immunosorbent assays. Selected sera were also tested by peptide immunoassays, by the four‐antigen recombinant immunoblot assay (RIBA II), and for viral genome by the polymerase chain reaction. Antibody to c100‐3 was detected in 61% of patients with chronic non‐A, non‐B (NANB) hepatitis and/or 46.5% with presumed NANB‐related cirrhosis by the first generation test. These figures increased to 77% and 58% when antibodies to recombinant structural and non‐structural HCV antigens were sought by the second generation assay. Supplemental testing against peptide Sp75 and Sp65isp67 confirmed that reactivity of sera by second generation assays was due to antibodies t o the additional structural and non‐structural antigens. Samples negative by the first generation assay were not confirmed by the supplemental assay using peptides Sp75 and Sp65/Sp67. HCV RNA was detected in 60% of the anti‐HCV positive sera tested, most of which were also RIBA II positive. Our findings confirm that the introduction of the structural and non‐structural antigens, especially the putativenucleocapsid protein, improves sensitivity of detection of antibodies to HCV, and facilitates diagnosis in patients with “cryptogenic” chronic hepatitis. © 1992 Wiley‐Liss, Inc.
Prevalence of herpesvirus, human T‐lymphotropic virus type 1, and treponemal infections in Southeast Asian refugees
Authors: Buchwald Dedra , Hooton Thomas M. , Ashley Rhoda L.
Published: March 2007

Sera obtained for treponemal serology (VDRL) from 193 Southeast Asian refugees representing five ethnic groups seen in a primary care clinic were examined for antibodies t o human T‐lymphotropic virus type 1 (HTLV‐1), human herpesvirus‐6 (HHV‐6), Epstein‐Barr virus (EBV), and cytomegalovirus (CMV). The seroprevalence was highest for EBV (99%), followed in decreasing order by CMV (95%), HHV‐6 (26%), and HTLV‐1 (0.6%). The VDRL was positive in 15% of patients. The highest seroprevalence to HHV‐6 was noted in the Chinese (33%) and the lowest in the Laotian hilltribes, the Mien and Hmong (14%). Antibody to HHV‐6 was most prevalent among patients under 20 and those between 60 and 69 years of age. Differences were not found among ethnic groups in the seroprevalence of HTLV‐1, EBV, or CMV. © 1992 Wiley‐Liss, Inc.
The detection in urine specimens of IgG and IgM antibodies to hepatitis A and hepatitis B core antigens
Authors: Perry Keith R. , Parry John V. , Vandervelde Elise M. , Mortimer Philip P.
Published: March 2007

The use of urine as a noninvasive specimen for the diagnosis of hepatitis A (HAV) and hepatitis B (HBV) virus infections was investigated. Specimens of urine were collected at the same time as blood or saliva specimens, or singly in cases of previously serologically confirmed recent infection. The specimens were tested for IgG and IgM anti‐HAV and anti‐HBc by immunoglobulin class‐specific capture radioimmunoassays (GACRIA and MACRIA). On the basis of assays on urine specimens it was possible to distinguish between individuals who were susceptible or immune to HAV or who had recently been infected with HAV. Using assays on 327 corresponding saliva specimens as reference tests, the observed sensitivity and specificity of tests on urine specimens by anti‐HAV GACRIA were 98.9% and 99.1%, respectively, and by anti‐HAV MACRIA were 95.8% and 99.6%, respectively. IgM and IgG anti‐HBc were detected readily in the urine of 35 acute or recent cases of hepatitis B but were not found in the urine of seronegative individuals. Of the urine specimens from 52 individuals who were HBsAg carriers or who had had long past HBV infections, 49 contained detectable IgG anti‐HBc. Of urine specimens from 42 HBsAg carriers, 11 contained raised IgM anti‐HBc levels.

Urine, which is a convenient specimen to collect, can be used t o study outbreaks of hepatitis A, to ascertain the HAV immune status of individuals, to differentiate hepatitis A from hepatitis B, and to identify individuals who have been naturally exposed to HBV. © 1992 Wiley‐Liss, Inc.
Mechanism of astrovirus entry into Graham 293 cells
Authors: Donelli Gianfranco , Superti Fabiana , Tinari Antonella , Marziano Maria Luisa
Published: March 2007

Astroviruses are intestinal pathogens associated with gastroenteritis in man and animals. The mechanism of internalization into host cells has not been reported previously. The cell entry pathway of serotype 1 human astrovirus into 293 cell line was studied biochemically and morphologically.

Viral infection was monitored by indirect immunofluorescence. Infected cells were treated with the lysosomotropic agents ammonium chloride, methylamine, and dansylcadaverine or the ionophore monensin to raise the intraendosomal and intralysosomal pH. All drugs tested inhibited the early stages of infection whereas they did not interfere with the viral binding to the plasma membrane.

The presence of astrovirus particles was detected by electron microscopy in coated pits and later in coated vesicles.

The data indicate adsorptive endocytosis as the most probable mechanism by which astroviruses enter susceptible cells. © 1992 Wiley‐Liss, Inc.
Hepatitis A vaccination in Greek military recruits
Authors: Papaevangelou G. , Totos Gr. , Roumeliotou A. , Safary A.
Published: March 2007

Improved standards of sanitation have contributed to a shift in the prevalence of hepatitis A in countries such as Greece. Children are now coming into first contact with the infection at an increasingly later age, leaving more adults susceptible to the disease. In military forces where close living conditions prevail, the likelihood of infection is even more pronounced. An inactivated hepatitis A vaccine has been developed and has been administered successfully to over 24,000 healthy children and adults. This vaccine would be of considerable benefit to military personnel worldwide.

The reactogenicity and immunogenicity of a hepatitis A vaccine were evaluated in 200 female military recruits, aged from 17 to 23 years, vaccinated according to a primary vaccination schedule at 0 and 1 months with a booster dose at 6 months.

Symptoms reported following vaccination were generally mild and transient. Soreness at the site of injection was the most frequent local symptom and malaise was the most common general symptom. Clinically significant increases in serum liver enzyme levels were not detected.

All subjects had seroconverted after the primary vaccination course and maintained anti‐HAV titres up to the time of the administration of the booster dose. The booster dose produced more than a tenfold increase in the geometric mean titre (GMT). © 1993 Wiley‐Liss, Inc.
Detection of neutralizing antibodies against human cytomegalovirus: Influence of strain variation
Authors: Weber B. , Klinghardt U. , Lux A. , Braun W. , Rabenau H. , Doerr H. W.
Published: March 2007

The influence of human cytomegalovirus (HCMV) strain variation on neutralizing antibody titers was investigated in sequential sera obtained from 12 organ transplant recipients (11 renal transplant recipients, 1 liver transplant patient) suffering from primary or secondary HCMV infection. Cross‐neutralization assays using either the international HCMV reference strain AD169 or individual clinical isolates from patients showed congruent results. Restriction enzyme analysis of the hypervariable α‐sequence of the L‐S junction of AD169 and HCMV isolates amplified by polymerase chain reaction (PCR) were carried out to confirm the expected strain differences. At least 6 groups of clinical strains were differentiated. The results of this study demonstrated that despite the strain heterogenicity of HCMV, the neutralization assay using AD169 permitted a reliable and quantitative serologic detection of neutralizing antibodies against HCMV. © 1993 Wiley‐Liss, Inc.
Changes in the prevalence of hepatitis B and C infection in patients with hepatocellular carcinoma in the Nagasaki prefecture, Japan
Authors: Hamasaki Keisuke , Nakata Keisuke , Tsutsumi Takuya , Tsuruta Shohtaro , Nakao Kazuhiko , Kato Yuji , Shima Masayoshi , Koji Toshihiko , Nagataki Shigenobu
Published: March 2007

The development of hepatocellular carcinoma (HCC) is very closely associated with chronic liver disease. In the present study, the prevalence of the hepatitis B virus (HBV) and hepatitis C virus (HCV) infection as a causative role in the development of HCC was analysed in 253 patients with HCC, who were admitted to our hospital during 1976–90. Among these patients, 68 (27%) were positive for HBsAg but negative for anti‐HCV antibody (group I); in contrast, 147 (58%) were negative for HBsAg but positive for anti‐HCV antibody (group II), 19 (7.5%) were both positive (group Ill), and 19 (7.5%) were both negative (group IV). To evaluate the serial changes in the prevalence of HBsAg and anti‐HCV antibody, changes in the number of patients were compared between group I and group II. The number of group I patients reached a peak during 1982–84 and was thereafter followed by a decreasing trend, whereas the number of group ll patients steadlly increased and reached a plateau over 6 recent years. These results suggest that HCV infection recently seems to play a more important role in the development of HCC than chronic HBV infection, even in the Nagasaki Prefecture, where the HBV carrier rate is higher than elsewhere in Japan. © 1993 Wiley‐Liss, Inc.
Lassa virus activity in Guinea: Distribution of human antiviral antibody defined using enzyme‐linked immunosorbent assay with recombinant antigen
Authors: Lukashevich I. S. , Clegg J. C. S. , Sidibe K.
Published: March 2007

More than 3,100 households in 27 selected villages distributed in the main geographic regions of Guinea were surveyed for the presence of Lassa virus‐specific IgG antibodies (LVA), using an enzyme‐linked immunosorbent assay (ELISA) with Lassa virus nucleocapsid protein expressed in insect cells infected with a recombinant baculovirus as antigen. The highest prevalence of LVA (25–55%) was found among inhabitants of tropical secondary forest (areas near Gueckedou, Yomou, and Lola) and guinea savannah (Faranah and Kindia areas), near the southern frontiers with Sierra Leone and Liberia. A much lower prevalence (4–7%) was found among inhabitants of mountainous (Pita, Labe, and Mali) and coastal (Boffa, Boké) areas. We found no discernible differences in LVA prevalence between males and females or among various age groups. Testing of 406 hospital staff members of the eight central hospitals in these areas for LVA revealed a similar distribution of seropositivity among hospitals in various prefectures. The highest prevalence of LVA in hospital staff (29–40%) was in the Gueckedou and Lola hospitals. Sera of LVA‐positive persons were tested via Western blot analysis. Antibodies bound predominantly to NP and GP2 proteins. © 1993 Wiley‐Liss, Inc.
Human parvovirus B19 infections: Routine diagnosis by a new nested polymerase chain reaction assay
Authors: Cassinotti Pascal , Weitz Manfred , Siegl Günter
Published: March 2007

A nested primer PCR assay was developed to detect human parvovirus B19 in various clinical specimens in a routine diagnostic laboratory. Under optimized conditions the highly specific assay had a sensitivity of less than 10 genome units. For practical reasons, however, this sensitivity was adjusted to 10–100 virus genomes in diagnostic applications. Using clinical specimens from 200 patients with suspected B19 infection, nested PCR was shown to have important diagnostic advantages over the detection of B19 specific antibodies. The data suggest that on the basis of serological data as obtained with currently available test systems a considerable proportion of B19 infections would be misdiagnosed. Examples for the usefulness of the PCR assay in routine diagnosis are given. © 1993 Wiley‐Liss, Inc.
Genetic variability of German hepatitis C virus isolates
Authors: Müller Hubert M. , Pfaff Eberhard , Goeser Tobias , Theilmann Lorenz
Published: March 2007

Heterogeneity of hepatitis C viral (HCV) genomes of several isolates from different countries has been reported, but there is little information on HCV isolates for the Federal Republic of Germany. Therefore, the nucleotide (nt) and deduced amino acid (aa) sequences of interesting parts of the viral genome derived from different human isolates in Germany were compared with each other and with the nt and predicted aa sequences of recently published isolates. HCV sequences were obtained by reverse transcription of viral RNA extracted from serum followed by polymerase chain reaction (PCR) amplification. Within the 5′ nontranslated region we found only 3 single nucleotide exchanges among 2 of our isolates, and in comparison to sequences of Japanese isolates 2 to 3 exchanges, and to U.S. isolates 1 to 5 exchanges (homologies 98% to >99%). Determination of a 249‐bp core sequence from two German isolates exhibited 3% sequence divergence. The sequence of the core region (nt 342–911) showed a homology of about 88–91% on nt level and 96–97% on aa level as compared to U.S. isolates and other German isolates, and a homology of 95–96% (nt) and 96–98% (aa), respectively, to Japanese isolates. Less homologies were noticed for the E1 and E2/NS1 genes, especially in the N‐terminal E2/NS1 hypervariable domain. Our isolates HD1 and HD2 showed nt sequence homologies of about 72–81% and aa homologies of 76–88% to U.S., German, and French isolates, and 89–91% (nt) and 89–96% (aa), respectively, to Japanese isolates. These results indicate that various German isolates are more closely related to Japanese isolates and differ from other European isolates as reported so far.

Because of a nucleotide sequence heterogeneity of up to 10% among the tested isolates, we conclude that more than one closely related but distinct viral genotype of HCV exists in Germany. Furthermore, heterogeneous sequences of HCV can be detected in a single patient suggesting multiple infection with different genomic variants or, alternatively, a genetic drift forced by mutational events as a consequence of host immune selection. © 1993 Wiley‐Liss, Inc.
Prevalence of canine distemper antibodies in the pagetic population
Authors: Gordon M. T. , Bell S. C. , Mee A. P. , Mercer S. , Carter S. D. , Sharpe P. T.
Published: March 2007

Recent molecular evidence has implicated canine distemper virus (CDV) as a possible aetiologic agent in Paget's disease. However, previous serological studies have shown no differences in levels of anti‐CDV antibodies between Paget's patients and controls.

In this study, the technique of enzyme linked immunosorbent assay was used to measure anti‐CDV antibodies in a group of Paget's patients from the North West of England. Some patients were undergoing treatment with 3‐amino‐hydroxypropylidene (APD), and the pre‐treatment levels of antibody were compared with those following treatment. With several patients, it was also possible to compare the antibody levels with results from in situ hybridisation studies.

No significant difference was found between the levels of anti‐CDV antibodies in patients and controls. However, several patients and some of the controls did have markedly elevated levels of anti‐CDV antibody. Antibody levels remained fairly constant following treatment with APD, except for two patients who showed marked changes.

The patients positive for CDV by in situ hybridisation had significantly lower levels of anti‐CDV antibodies when compared with those that were negative by in situ hybridisation.

These results suggest that if CDV does cause Paget's disease, anti‐CDV antibodies play little or no part in the disease pathogenesis. High levels of anti‐CDV antibodies in both Paget's patients and controls suggest that a canine virus can infect humans. The fact that those patients that had CDV transcripts detectable in their bone cells had low levels of anti‐CDV antibodies might suggest failure to clear the virus during an initial infection. This might then allow sequestration of CDV within bone cells and, thus, eventually lead to Paget's disease. © 1993 Wiley‐Liss, Inc.
Hepatitis C antibodies in dialysis patients and patients with leukemia
Authors: Kolho E. , Oksanen K. , Honkanen E. , Naukkarinen R. , Krusius T.
Published: March 2007

The prevalence of hepatitis C antibodies (anti‐ HCV) among multitransfused patients was studied and compared with predicted values obtained from a post‐transfusion hepatitis study and from data on the prevalence of anti‐HCV among blood donors. The prevalence of hepatitis B core antibodies (anti‐HBc) was also studied to determine the routes of transmission of hepatitis C virus. The patients consisted of 65 dialysis patients (57 on haemodialysis and 8 on continuous ambulatory peritoneal dialysis) and 71 leukemia patients in long‐term remission [49 with acute myeloid leukemia (AML) and 22 with acute lymphatic leukemia (ALL)]. The presence of anti‐HCV was investigated using a second generation enzyme‐linked immunosorbent assay. Reactive samples were confirmed by a second generation recombinant immunoblot assay. Anti‐HBc was studied in the 65 dialysis patients and in 40 of the leukemia patients. Three (4.6%) of the 65 dialysis patients and 12 (24.5%) of the 49 AML patients were anti‐HCV positive whereas all of the ALL patients were seronegative. The total number of blood units transfused to 134 patients (data on two dialysis patients were not available) was 18,148, out of which 17,575 units had been transfused prior to the initiation of anti‐ HCV screening of blood donors. On the basis of the anti‐HCV prevalence among blood donors and the incidence of post‐transfusion hepatitis, the predicted number of seropositive patients was 11 and 18, respectively. Five of the 65 dialysis patients were anti‐HBc positive, compared with only one of the 40 leukemia patients. It is concluded that the anti‐HCV prevalence among dialysis and leukemia patients is concordant with the risk of receiving contaminated blood products, whereas hepatitis B infection may have other routes of transmission in dialysis patients. © 1993 Wiley‐Liss, Inc.
Comparative analysis of the restriction endonuclease profiles of the Dumas and Singapore strains of varicella‐zoster virus
Authors: Chow Vincent T. K. , Wan Simon S. Y. , Doraisingham S. , Ling A. E.
Published: March 2007

The incidence of varicella in Singapore has been increasing since 1984. In 1991,17,930 cases were reported in a population of about 3 million. A serological survey completed in 1990 demonstrated that only 43% of the cohort had antibodies to varicella‐zoster virus (VZV), indicating inadequate herd immunity. To exclude novel VZV strains, representative VZV isolates from 9 chicken pox and 4 zoster patients were characterised by restriction endonuclease analysis. DNAs were extracted from viral isolates propagated in MRC5 human embryo lung cells and were digested separately with Bg/ll, EcoRI, Pstl, Sall, and Xbal enzymes. The cleavage profiles of these VZV strains derived from both chicken pox and zoster lesions revealed no distinct differences. This observation implies that the current upsurge of chicken pox most likely stems from closely related VZV genotypes infecting a susceptible population with insufficient herd immunity. Comparison of the restriction fragments of the Singapore and the Dumas strains revealed polymorphisms of the Sal/I‐D, SaI/l‐E, and Xbal‐l fragment lengths, which correlated with variable regions I, II, and Ill of the VZV genome, thereby representing geographically distinct genotypic variants of VZV. © 1993 Wiley‐Liss, Inc.
Frequent isolation of human herpesvirus 7 from saliva samples
Authors: Hidaka Yasufumi , Liu Ying , Yamamoto Masahiro , Mori Ryoichi , Miyazaki Chiaki , Kusuhara Koichi , Okada Kenji , Ueda Kohji
Published: March 2007

Human herpesvirus 7 (HHV‐7) was isolated frequently from saliva specimens. The isolation rates were 81% (13/16) in adults, 70% (7/10) in children over 1 year old, and none (0/7) in children less than 1 year old, respectively, indicating that infection of HHV‐7 occurs during early infancy and the virus shedding rate after infection is very high. Human herpesvirus 6 (HHV‐6) was not isolated from saliva specimens although some studies on isolation of HHV‐6 from saliva was reported previously. © 1993 Wiley‐Liss, Inc.
Immunogenicity of 20 μg of recombinant DNA hepatitis B vaccine in healthy neonates: A comparison of three different vaccination schemes
Authors: Canho R. Del , Grosheide P. M. , Voogd M. , Huisman W. M. , Heijtink R. A. , Schalm S. W.
Published: March 2007

The immunogenicity of a full dose (20 μg) of recombinant DNA yeast‐derived hepatitis B vaccine (Engerix‐B) was assessed in healthy neonates in order to compare three candidate vaccination schemes. After randomization 162 newborns of hepatitis B surface antigen (HBsAg) negative mothers entered the study. Neonates received hepatitis B vaccine according to a fourdose vaccination scheme starting either at month 3 (scheme I: months 3,4,5, and 11) or at birth (scheme III: months 0,1,2, and 11). Another group of neonates received hepatitis B vaccine according to a three‐dose scheme starting at birth (scheme II: months 0, 1, and 6). Serious adverse reactions were not observed; 2.5% of the vaccinated newborns suffered mild transient local symptoms. The vaccine was highly immunogenic irrespective of vaccination scheme; all infants developed anti‐HBs levels ≥10 IU/L, 97% ≥100 IU/L. The immunogenicity of hepatitis B vaccine after primary and booster vaccinations, administered in the four‐dose scheme started at birth, was significantly higher (P< 0.05) than in the three‐dose scheme started at birth. Hepatitis B vaccination according to the four‐dose scheme started at month 3 produced significantly higher (P < 0.05) antibody levels in comparison to the four‐dose scheme started directly after birth. This study showed that a fourdose hepatitis B vaccination scheme starting at month 3 resulted in the highest antibody levels of the three schemes investigated and can be recommended for incorporation in the Expanded Programme on Immunization in The Netherlands.
Second generation hepatitis C virus assays: Performance when testing african sera
Authors: Callahan Johnny D. , Constantine Niel T. , Kataaha Peter , Zhang Xiang , Hyams Kenneth C. , Bansal Jaya
Published: March 2007

Sera were collected from 426 volunteers in Uganda at high and low risk for acquisition of hepatitis C virus (HCV). All samples were tested by the Ortho HCV second generation ELISA (S1) and by the INNOTEST HCV Ab second generation enzyme immunoassay, (S2), (Innogenetics, Antwerpen, Belgium). Sera that were repeatedly reactive by either screening assay were further tested by each of two different HCV supplemental/confirmatory assays: a second generation recombinant immunoblot assay (RIBA, Ortho Diagnostics), (C1), and a line immunoassay (INNO‐LIA HCV‐Ab, Innogenetics), (C2).

In these populations there were 16 true positives, 351 true negatives, and 59 indeterminate results. Fifty‐nine point four percent (253/426) of the samples were repeatedly reactive by the S1 test, while only 2.6% (11/426) were repeatedly reactive by S2. Test S1 produced a high false positive rate, a low positive predictive value, a specificity of 49.3%, and had a sensitivity of 100%. In contrast, the S2 screening assay had much higher specificity (98.8%), but lacked in sensitivity (31.3%). This poor sensitivity of S2 was based almost exclusively on the fact that the C2 supplemental test classified 9 samples as confirmed positive when the homologous screening assay classified these samples as negative; several of these were not confirmed when using a new generation INNO‐LIA. Both the screening tests S1 and S2, and the supplemental assays C1 and C2 exhibited a significant degree of discordance, and neither of the screening tests alone would be adequate for use in these populations.
Human papillomavirus type 16 variants isolated from vulvar Bowenoid papulosis
Authors: Fang Bruno Schwartzman , Guedes Antonio Carlos , Muñoz Luiz Carlos , Villa Luisa Lina
Published: March 2007

Tissues from two cases of Bowenoid papulosis of the vulva were found to contain human papillomavirus (HPV) 16 DNA by Southern blot hybridization. Analysis of the hybridization pattern revealed differences in a restriction fragment of one specimen as compared to the HPV 16 DNA prototype. To investigate if these differences could interfere with the expression of such oncogenic viral genomes, the corresponding DNA fragments were cloned and further analyzed. After amplification by PCR and DNA sequencing, a 213 base pairs duplication was mapped in the long control region (LCR) of this HPV 16 variant. One single PCR fragment was obtained from the other Bowenoid papulosis, which is identical in size with the same region in the HPV‐16 prototype. The duplication in the HPV‐16 LCR analyzed in this study maps upstream of a region containing several regulatory elements.
Administration of inactivated and detergent‐treated influenza virus to mice before virus challenge reduces mortality
Authors: Arora D. J. S.
Published: March 2007

Formalin‐inactivated virus (FIV) and the detergent‐treated virus (DTV) preparations were tested for their ability to enhance the resistance of mice to experimental influenza infection. FIV (100 μg) was administered intravenously to mice. After 24 hr, animals were challenged with 5 LD50 dose intranasally. FIV‐treated and nontreated (control) mice had 10% and 100% mortality, respectively. Similar results were obtained with the DTV (40 μg) preparation. The pulmonary virus titer of FIV‐treated mice was lower when compared with the control. Mechanisms other than acquired immunity may have conferred the early resistance to virus infection in mice.
Demonstration of hepatitis C virus genome in saliva and urine of patients with type C hepatitis: Usefulness of the single round polymerase chain reaction method for detection of the HCV genome
Authors: Numata Noboru , Ohori Hitoshi , Hayakawa Yasuhiko , Saitoh Yukie , Tsunoda Akira , Kanno Atsushi
Published: March 2007

The possible transmission routes of hepatitis C virus (HCV) in patients without overt parenteral exposure (sporadic or community acquired form) were examined. Saliva and urine specimens obtained from type C hepatitis patients, whose sera were positive for the HCV genome, were examined by reverse transcription and polymerase chain reaction (RT‐PCR). By analyzing the factors that influenced the detection of the HCV genome by PCR, we developed a single round method which enabled semiquantitative detection with higher sensitivity than that obtained with nested PCR. Single round PCR revealed that 34.8% (8 of 23) of saliva and 56.5% (13 of 23) of urine specimens from patients with type C hepatitis contained the HCV genome. The amounts of HCV genome in saliva and urine specimens correlated with those in serum. The relative amounts of HCV genome in serum, saliva, and urine from a chronic type C hepatitis patient were determined by comparing the reciprocal of the smallest volume of the specimens in which the PCR products were visualized in agarose gels (PCR units/ml), and the values were 1 ×105, ×101, and 3 ×101 PCR units/ml for serum, saliva, and urine specimens, respectively.
Quantitative analysis of herpes simplex virus DNA in cerebrospinal fluid of children with herpes simplex encephalitis
Authors: Ando Yoshihiro , Kimura Hiroshi , Miwata Hiroyuki , Kudo Toyoichiro , Shibata Motohiro , Morishima Tsuneo
Published: March 2007

Herpes simplex virus (HSV) DNA in the cerebrospinal fluid (CSF) of children with herpes simplex encephalitis (HSE) was quantified and typed using the polymerase chain reaction (PCR) assay. During the acute phase, HSV‐DNA was detected in the CSF of 13 patients with HSE, including 5 neonates. A restriction profile of the PCR products cleaved with the restriction enzymes Xhol and Bglll showed that 2 neonatal samples were HSV‐2, and the remainder were HSV‐1. The amount of HSV‐DNA in the initial CSF ranged from 102105 copies/ml. A significantly greater number of HSV‐DNA copies was detected in neonates than in older children (mean 3.9 vs. 2.5, log10 copies/ml p < .05). Except for one patient, the amount of HSV‐DNA decreased gradually with acyclovir therapy. These results show that a quantitative PCR assay is applicable not only to the diagnosis of HSE but also for monitoring the response to antiviral drugs.
Confirmation of hepatitis C virus antibody in blood donors
Authors: Dow B. C. , Coote I. , Munro H. , McOmish F. , Yap P. L. , Simmonds P. , Follett E. A. C.
Published: March 2007

Of 103,203 donations collected in Scotland and Northern Ireland over a 3‐month period and screened for HCV antibody by Ortho or Abbott second‐generation ELISAs, 340 were found repeatedly reactive. Supplementary testing with RIBA‐2 resulted in 77 being classified as positive, 130 as indeterminate, and 133 as negative. PCR analysis of the positives and indeterminates indicated viraemia in 65 (84%) of the positives and 7 (5.5%) of the indeterminates. To determine if PCR analysis could be eliminated or reduced by further serological testing, all RIBA‐2 positives and indeterminates were tested by UBI and Wellcozyme ELISAs and Innolia and RIBA‐3 immunoblots. All RIBA‐2 positives with bands to more than 1 gene product were detected in all 4 systems, but >60% of RIBA‐2 indeterminates were negative in those tests that contain either recombinant antigens or synthetic peptides derived independently from those used by Ortho/Abbott tests. A comparison of data from the 79 reactive with the core (c22) region revealed only 16 samples reactive in all 4 systems as well as Ortho and Abbott. These 16 included all 6 of the PCR positives in the 79 c22 indeterminate samples. ELISAs and immunoblots using independently derived antigens can offer a useful method of screening out nonspecific reactions in Ortho or Abbott ELISAs, hence reducing the need for PCR testing. Some caution is required as all such tests do not contain identical mixes of antigenic material.
Characterization of minor and major antigenic regions within the hepatitis B virus nucleocapsid
Authors: Tordjeman Marc , Fontan Gilles , Rabillon Véronique , Martin Jacques , Trepo Christian , Hoffenbach Agnés , Mabrouk Kamel , Sabatier Jean Marc , van Rietschoten Jurphaas , Somme Gérard
Published: March 2007

Hepatitis B core antibodies (anti‐HBc) appear very early during the course of the hepatitis B virus infection and often persist years after viral clearance. In order to characterize the immunodominant domain of the HBcAg, the human immune response against the HBV nucleocapsid (HBcAg) was analyzed by using 14 synthetic peptides. Anti‐HBc antibodies were detected by an indirect enzyme‐linked immunosorbent assay (ELISA) with HBc peptides. Results suggest that the anti‐HBc response is heterogeneous and directed against the whole primary structure of the HBc protein. Results also indicate that the epitopes recognized by anti‐HBc antibodies can vary with the stages of the disease. In most sera from patients with serological evidence of acute HBV infection, anti‐HBc antibodies recognized all the HBc peptides; conversely, after the acute phase, anti‐HBc antibodies recognized predominantly epitopes located within the central region of the HBc protein from residue 74 to 123. Our results suggest that the HBV core protein is made up of two antigenic regions: a major one expressing a family of immunodominant epitopes from residue 74 to 123, whereas the minor encompasses the rest of the protein. The concept of the conformational nature of the unique HBcAg determinant is discussed, suggesting numerous families of linear epitopes.
Partial sequence comparison of eight new Chinese strains of hepatitis E virus suggests the genome sequence is relatively stable
Authors: Yin Shurong , Tsarev Sergei A. , Purcell Robert H. , Emerson Suzanne U.
Published: March 2007

Partial genomic sequences representing 420 nucleotides of a nonstructional region, 480 nucleotides of the putative RNA polymerase region, and 540 nucleotides of the structural region of epidemic‐associated Chinese strains of hepatitis E virus (HEV) were obtained by direct sequencing of PCR‐amplified DNA. Comparison with previously published HEV sequences showed a clear relatedness of all Chinese strains to each other and to a Pakistani strain (Sar‐55). All eight Chinese strains examined had very similar sequences (98.5‐99.8% homology) in the regions examined and were much closer to the Pakistani strain (Sar‐55) (97.9‐98.4% homology) than to the Burmese strain (92.5‐93.3% homology). Sequence comparisons of the three genomic regions in the Chinese strains indicated that the RNA polymerase region was much more conserved than the other nonstructural region or the structural region. HEV isolates from three remote geographic regions of China had sequences closely related to each other.
Characterization of Epstein‐Barr viral strains in parotid gland saliva and peripheral blood of patients with primary Sjögren's syndrome and healthy EBV carriers
Authors: Oosterveer M. A. P. , Markusse H. M. , Lennette E. T. , Zou J. Z. , Bolhuis R. L. H. , Gratama J. W.
Published: March 2007

Increased Epstein‐Barr virus (EBV) replication has been reported in the salivary and lacrimal glands in Sjögren's syndrome (SS). We studied whether or not certain EBV strains would occur preferentially in the peripheral blood and parotid gland saliva of 18 EBV‐seropositive patients with primary Sjögren's syndrome (pSS) and 12 EBV‐seropositive control persons. Transforming EBV was detected in the blood of 11 of 18 (61%) pSS patients and 9 of 12 controls (75%). Unexpectedly, neither transforming nor Raji‐superinfecting EBV strains were detected in SS parotid saliva, whereas these EBV types were detected in control saliva in 7 and 8 cases, respectively (P <0.001). Transforming EBV strains were further characterized by ‘Ebnotyping,’ i.e., analysis of the size spectrum of the viral antigens EBNA 1, 2, 3, and 6 in immunoblots of lymphoblastoid cell lines (LCL). Previous work has shown that a single EBV strain (Ebnotype) dominates the blood and oropharynx of healthy carriers and that unrelated individuals carry different EBV strains, reflecting the vast polymorphism of Ebnotypes in the general population. Two unexpected observations were made. First, an identical Ebnotype was detected in 4 unrelated individuals, i.e., in the blood of 1 pSS patient and in the saliva of 3 control persons. Second, carriage of 2 to 4 different Ebnotypes by a single individual was observed in 4 cases, i.e., in the blood of 1 pSS patient, and in the blood and saliva of 3 control persons. As only 1 control person had received a blood transfusion, it is suggested that (super)infection with exogenous EBV strains via salivary transmission and/or recombination of endogenous virions may have contributed to this situation.
BK virus infection of the human urinary tract
Authors: Shinohara Toshiya , Matsuda Michiyuki , Cheng Seng H. , Marshall John , Fujita Miri , Nagashima Kazuo
Published: March 2007

By screening consecutive autopsy cases with an antibody that recognizes human polyomaviruses, we found a case of malignant lymphoma in which the virus infection was confined to epithelia of the renal calyces, renal pelvis, ureter, and urinary bladder. The virus was confirmed as BK virus by a specific monoclonal antibody against BK virus T antigen, and numerous virus particles were identified by electron microscopy. The results showed that BK virus is a human urotheliotrophic virus.
Outbreak of adenovirus type 4 conjunctivitis in South Australia
Authors: Schepetiuk Sonia K. , Norton Robert , Kok Tuckweng , Irving Louise G.
Published: March 2007

An outbreak of adenovirus type 4 conjunctivitis occurred in South Australia between April and November 1992. Eye swabs were submitted by general practitioners and ophthalmologists who had seen patients with clinical conjunctivitis or keratitis. Apart from interfamilial spread, there were no other common epidemiological factors. Adenovirus was isolated from the eye swabs of 38 patients. Isolates were typed by neutralisation tests and restriction endonuclease cleavage patterns and found to be adenovirus type 4. This report serves to illustrate an infrequent cause of epidemic conjunctivitis, namely adenovirus type 4. There was no demonstrable focus of the outbreak.
Immune recognition of linear antigenic regions within the hepatitis B pre‐C and c‐gene translation products using synthetic peptides
Authors: Sällberg Matti , Rudén Ulla , Wahren Britta , Magnius Lars O.
Published: March 2007

The antibody recognition of linear regions within the amino acid (aa) sequence of hepatitis B (HB) core antigen (HBcAg), e antigen (HBeAg), and pre‐C region was investigated in 46 patients infected with hepatitis B virus (HBV), and one immunized rabbit. Peptide analogues were synthesized to cover the complete product of the C‐gene, including the pre‐C region using various synthetic methods. Two carriers of hepatitis B surface antigen (HBsAg) with anti‐HBe, recognized pin‐bound decapeptides covereing amino acid (aa) 76‐83 of HBc/eAg, and the most essential residues were found to be Asp78, Pro79, Arg82, and Asp83. Pre‐C peptides were recognized by lgG1 or lgG3 in sera from two out of ten cases with acute HB, in four out of twelve sera from HBeAg‐positive carriers of HBsAg, and in two out of twelve sera from anti‐HBe‐positive carriers of HBsAg. Two sera from the cases of acute HB showed strong reactivity of the IgG3 isotype with HBc/eAg peptides 61–85. Five of the sera from HBeAg‐positive carriers of HBsAg were weakly reactive with peptides 41–60, 61–85, 121–140, and/or 141–160. Eight of the sera from anti‐HBe‐positive carriers of HBsAg recognized aa 121‐140 of HBc/e with lgG1, lgG3, and/or lgG4 iso‐types. IgG from one immunized rabbit recognized peptides 1–20, 61–85, and 71–90, and the T‐cells recognized peptides 1–20 and 71–90. Thus, human and rabbit antibodies recognize linear antigenic regions within the pre‐C, and within regions 1–20, 41–60, 61–85, 121–140, and 141–160 of HBcAg. Thus, the major HBc site is likely to comprise more than one part of the HBcAg sequence, of which a sequence within residues 61–85 might be a linear component. © 1994 Wiiey‐Liss, Inc.
Genome type analysis of chilean adenovirus strains isolated in a children's hospital between 1988 and 1990
Authors: Kajon Adriana E. , Larrañaga Carmen , Suárez Mónica , Wadell Göran , Avendaño Luis F.
Published: March 2007

In a study designed to evaluate the genetic variability of adenovirus strains associated with infantile cases of respiratory disease requiring hospitalization, a collection of 136 adenovirus isolates obtained in the Roberto del Rio Children's Hospital of Santiago, Chile between June 1988 and November 1990 was studied by restriction enzyme analysis. Nasopharyngeal aspirates were obtained on admission from children under 2 years. During the study period a total of 227 adenovirus respiratory infections (ARI) were diagnosed at the ward for ARI by immunofluorescence, representing 23% of all admissions. Fifty percent of the 136 typed strains were found to belong to subgenus B, and the other 50% corresponded to subgenus C. Digestion with a set of seven enzymes allowed the identification of nine different genome types of subgenus C, three of which had not been previously described, exhibiting novel restriction patterns with either Bgl II or BstEII. Ad7h, identified in 66 isolates, was the predominant genome type and was associated with the nine cases requiring mechanical respiratory assistance and with the two fatalities recorded during the 29 months. No differences were found between the age and sex distribution of subgenus B and C genomic variants, but the mean length of hospital stay (X ± 2 SE) recorded among patients infected with subgenus B types was significantly higher (17.72 + 4.52 days (n = 55) vs. 7.54 + 1.70 days (n = 53); F = 17.22; P < 0.0001) © 1994 Wiiey‐Liss, Inc.
High titers of anti‐epstein‐barr virus DNA polymerase are found in patients with severe fatiguing illness
Authors: Natelson Benjamin H. , Ye Ning , Moul Douglas E. , Jenkins Frank J. , Oren Dan A. , Tapp Walter N. , Cheng Yung‐Chi
Published: March 2007

Forty‐one patients with chronic fatigue syndrome (CFS), 76 healthy controls matched with the patient group for age range, sex, race, and socioeconomic class, and 22 symptomatic patients with seasonal affective disorder (SAD) had serum sampled for antibodies against 2 Epstein‐Barr virus (EBV) replicating enzymes. Abnormal liters of antibodies were found twice as often in CFS patients as controls (34.1% vs. 17.1%), with SAD patients having an intermediate frequency (27.3%). Stratifying for disease severity sharpened the differences considerably, with the sicker CFS and SAD patients having 52% and 50% abnormal tests, respectively; more mildly afflicted CFS and SAD patients had a frequency of abnormal tests in the normal range. Antibodies to EBV DNA polymerase (DNAP) were the more sensitive of the two tests in that they were positive in all cases but one. These findings suggest that antibodies against EBV DNAP may be a useful marker in delineating a subset of patients with severe fatiguing illness for appropriate treatment trials and for monitoring their outcomes. © 1994 Wiiey‐Liss, Inc.