bluuebunny/crossref_metadata_2025 huggingface dataset:
GACPAT HIV 1+2: A simple, inexpensive assay to screen for, and discriminate between, anti‐HIV 1 and anti‐HIV 2
Authors: Parry John V. , Connell Jeffrey A. , Reinbott Paul , Mortimer Philip P. , Garcia Ana B. , Avillez Francisca
Published: March 2007

A simple and cheap assay suitable for screening for anti‐HIV 1 and anti‐HIV 2 and discriminating between them was evaluated. In it specimens are incubated in U‐bottomed microplate wells coated with anti‐human IgG for 30 min at room temperature. After washing, 100 μl of a 1 in 50 dilution of HIV 1‐coated gelatin particles (Serodia‐HIV 1/2, Fujirebio) are added. Settling patterns are read on the second day: A positive reaction is indicated by adherence of the particles and a negative by a button. The HIV 1 particles are then washed away and HIV 2 particles added. Anti‐HIV 2 reaction patterns are read on the third day. To assess the performance of the modified “GACPAT HIV 1+2” assay a panel of 1,621 serum/plasma specimens was used. It comprised validated anti‐HIV 1 positive (n = 220), anti‐HIV 2 positive (n = 214), dual anti‐HIV 1/anti‐HIV 2 positive (n = 11), and anti‐HIV negative (n = 1,176) serum/plasma specimens.

All 434 specimens that contained anti‐HIV 1 or anti‐HIV 2 reacted positively with the homologous particles. The 11 dually positive specimens reacted positively with both HIV 1 and HIV 2 particles. Five (2.3%) anti‐HIV 1 and five (2.3%) anti‐HIV 2 positive specimens gave positive reactions with both particle types, but none of the five cross‐reactive anti‐HIV 2 specimens were dually reactive when the order of particle addition was reversed. One repeat false positive reaction was recorded with the HIV 1 particles and none with the HIV 2 particles, giving specificities of 99.9% and 100%, respectively. Type specificity was 98.8% after the results of the reverse assay had been taken into account. The assay was sensitive for anti‐HIV 1 in seroconversion series and for anti‐HIV 2 in highly diluted specimens.

GACPAT HIV 1 +2 is an accurate anti‐HIV assay applicable to serum/plasma. With few exceptions anti‐HIV 1 positive specimens are reactive only on the second day and anti‐HIV 2 positive specimens only on the third day. It thus discriminates anti‐HIV 2 from anti‐HIV 1, probably through depletion of cross‐reacting antibodies by the reaction with the particles initially added. © 1995 Wiley‐Liss, Inc.
Is the detection of anti‐hepatitis C virus core IgM influenced by the presence of serum rheumatoid factor?
Authors: Pawlotsky Jean‐Michel , Remire Jocelyne , Darthuy Françoise , Intrator Liliane , Udin Laure , Dhumeaux Daniel , Duval Jean
Published: March 2007

Rheumatoid factor (RF) induces false‐positive results in the detection of serum antibodies, especially of the IgM type. About 70% of the patients with chronic hepatitis C have abnormal levels of serum RF. The aim of this study was to determine whether the presence of serum RF could influence the detection of anti‐HCV core IgM, using an assay designed not to pick up RFs by the addition of goat antibodies directed against human IgG in the sample diluent. Serum anti‐HCV core IgM antibodies and RF were sought in 60 patients with chronic hepatitis C. Serum anti‐HCV IgG antibodies and anti‐HCV core IgM antibodies were also sought in 101 patients with high levels of RF. Anti‐HCV core IgM antibodies were found in 45% and serum RF in 72% of the patients with chronic hepatitis C. Neither the prevalence nor the levels of RF differed significantly between IgM positive and negative patients. Eight percent of the 101 patients with raised RF had anti‐HCV antibodies and two of them had anti‐HCV core IgM antibodies. No patient without anti‐HCV antibodies had anti‐HCV core IgM antibodies. These results show that: a) the detection of anti‐HCV core IgM in patients with chronic hepatitis C is independent of the presence of serum RF; b) high titers of serum RF are not responsible for false‐positive results of anti‐HCV IgM tests. The study suggests that the test used could be a confident tool for studies on the significance of anti‐HCV core IgM antibodies in chronic hepatitis C. © 1995 wiley‐Liss, inc.
Infectious virus titer, replicative and syncytium‐inducing capacity of human immunodeficiency virus type 1
Authors: Nkengasong John N. , Peeters Martine , Nys Patrick , Willems Betty , Piot Peter , van der Groen Guido
Published: March 2007

The relationship was investigated between a viral infectious titer in peripheral blood mononu‐clear cells (PBMC) and plasma on the replicative and syncytium‐inducing capacity of human immunodeficiency virus type 1 (HIV‐1) isolates. The replicative capacity was defined as the minimum time required for p24 antigen to become positive in PBMCs or plasma of HIV‐1 infected individuals, cocultured with PBMCs of healthy donors. Syncytium induction was determined by the MT‐2 cell assay and defined as the presence of giant multinucleated cells. The replicative capacity of HIV‐1 in PBMCs of healthy donors correlated with the infectious viral titer in PBMCs, but not in the plasma of HIV‐1 positive patients. Syn‐cytia formation in MT‐2 cells was not related to the infectious viral titer in PBMCs or plasma of HIV‐1 positive patients. These findings suggest that syncytium formation, not replicative capacity, is an intrinsic HIV‐1 phenotype. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus RNA in plasma and blood mononuclear cells in patients with chronic hepatitis C treated with alpha‐interferon
Authors: Ounanian A. , Seigneurin J. M. , Gueddah N. , Rolachon A. , Thelu M. A. , Zarski J. P.
Published: March 2007

Hepatitis C virus (HCV), a positive stranded RNA virus, is the main causative agent of post‐transfusion and sporadic non‐A non‐B hepatitis worldwide. Paired samples of plasma and peripheral blood mononuclear cells (PBMC) from 11 patients with chronic hepatitis C treated with α‐interferon (IFN) were tested, using a single step polymerase chain reaction (PCR), for the presence of HCV RNA. Before treatment, the viral genome was detected in all the plasma samples and 81.8% of PBMC. After 3 months of treatment, HCV RNA was still present in 63.6% of plasma samples but in only 27.3% of PBMC. A good correlation was observed between serum alanine aminotransferase level normalisation and disappearance of the viral genome in plasma. Among the six responder patients, five relapsed shortly after IFN withdrawal; HCV RNA became detectable again, especially in PBMC. These results show the presence of HCV in PBMC from most patients infected chronically. IFN therapy had an inhibitory effect on viral replication in lymphoid cells, but frequent relapses observed after cessation of treatment with IFN suggested persistence Of HCV in these Cells. © 1995 Wiley‐Liss, Inc.
Comparison of the kinetics of puumala virus specific IgM and IgG antibody responses in nephropathia epidemica as measured by a recombinant antigen‐based enzyme‐linked immunosorbent assay and an immunofluorescence test
Authors: Elgh Fredrik , Wadell Göran , Juto Per
Published: March 2007

Immunoglobulin M and G (IgM and IgG) responses were followed up to 6 months in patients with nephropathia epidemica (NE) by an enzyme‐linked immunosorbent assay (ELISA) using a recombinant Puumala virus (PUU) nucleocapsid protein as antigen and an immunofluorescence test (IF) using PUU infected, acetone‐treated cells as antigen. The recombinant protein was produced by cloning and expressing the nucleocapsid encoding gene of PUU as a polyhistidine fusion protein in Escherichia coli. The product was purified over a metal chelating ion affinity column. On admission, all 17 patients had an IgM response by both methods. The IgM titers decreased significantly by both methods 3 months after onset (ELISA P<0.05 and IF P<0.05). Four of six still had detectable IgM, however at low levels, after 6 months. Presence of specific IgG differed significantly on admission between the two methods: by ELISA 8 of 17 had detectable specific IgG, whereas by IF 15 of 17 had specific IgG (P < 0.02). There were 10 significant titer rises between acute and convalescent serum samples in the same patients by both methods. It is concluded that the IgG antibody response differs in the early phase of NE as measured by a method using a recombinant PUU nucleocapsid protein and a method using PUU infected acetone‐treated cells as antigens. Furthermore, the results suggest that it is of importance to rely on specific IgM for serodiagnosis of NE during the acute phase. © 1995 Wiley‐Liss, inc.
Clinical characteristics and antibody profiles of chronic hepatitis C patients: Relation to hepatitis C virus genotypes
Authors: Yuki Nobukazu , Hayashi Norio , Mita Eiji , Hagiwara Hideki , Oshita Masahide , Ohkawa Kazuyoshi , Katayama Kazuhiro , Kasahara Akinori , Fusamoto Hideyuki , Kamada Takenobu
Published: March 2007

Genotyping of 179 consecutive Japanese chronic hepatitis C patients was carried out based on the variation in the hepatitis C virus (HCV) core gene. The results were correlated with clinical features and antibody responses toward specific HCV proteins deduced from the nucleotide sequence of genotype I/1 a. Genotypes II/1 b, III/2a, and IV/2b were identified in 138 (77%), 24 (13%), and 12 (7%) patients, respectively. Five patients had double infections. Genotype dependence was observed only for antibody response toward the NS4 (5–1–1) protein, which was infrequent in genotype III/2a patients (33%) compared with genotype II/1 b (81%; P < 0.01) and genotype IV/2b (75%; P < 0.05). Following interferon‐α therapy, sustained aminotransferase normalisation was achieved by 89% (eight of nine) patients without antibody to the 5–1–1 protein and 33% (17 of 51) with it (P < 0.01). These findings indicate that absence of antibody response to the 5–1–1 protein is frequent in genotype III/2a HCV carriers and may serve to predict responses to interferon therapy. © 1995 Wiley‐Liss, Inc.
Renal failure due to BK virus infection in an immunodeficient child
Authors: De Silva L. M. , Bale P. , De Courcy J. , Brown D. , Knowles W.
Published: March 2007

We report a child with cartilage‐hair hypoplasia and Hodgkin's disease who developed progressive renal failure and died following infection with a polyomavirus, BK virus. Renal biopsy showed interstitial inflammation, tubular atrophy, and intranuclear inclusions in tubular epithelium, with large numbers of papovavirus particles within the cells. BK virus infection was confirmed by polymerase chain reaction on renal biopsy material and in urine and the demonstration of a high titre of haemagglutination inhibition (HI) and IgM antibody to the virus in the patient's serum. This case emphasises the need to study in depth any unusual clinical manifestation in immunocompromised patients to delineate better the full clinical impact of less well‐established pathogens such as BK virus. © 1995 Wiley‐Liss, Inc.
Polymerase chain reaction detection of small round‐structured viruses from two related hospital outbreaks of gastroenteritis using inosine‐containing primers
Authors: Green Steve M. , Lambden Paul R. , Deng Yu , Clarke Ian N. , Lowes J. Andrew , Lineham Sarah , Bushell Julie , Rogers John , Caul E. Owen , Ashley Charles R.
Published: March 2007

Two outbreaks of gastroenteritis in the UK which occurred nine days apart at Lymington and Southampton hospitals were investigated. The clinical and epidemiological features of both outbreaks were characteristic of small round‐structured virus (SRSV) infection with rapid onset of diarrhoea and/or nausea and vomiting and propagation of the outbreaks by secondary spread. SRSV particles were observed by immune electron microscopy (EM) in 60% of faecal samples from both outbreaks and no other pathogens were detected. The index case for the second outbreak was a patient who was admitted with diarrhoea and vomiting after being discharged from Lymington hospital during the first outbreak. The possibility that the two outbreaks were caused by the same strain of SRSV was investigated by the polymerase chain reaction (PCR). New inosine‐containing PCR primers were designed to amplify the RNA polymerase region of SRSV cDNA from genetic groups I and II. The PCR using the group II primers achieved a higher detection rate for SRSVs in faecal samples (68% of samples positive from both outbreaks) than immune EM. SRSVs were not detected using the group I primers or using conventional degenerate PCR primets. The nucleotide sequences of PCR amplicons from both outbreaks were identical providing molecular epidemiological evidence for the involvement of a single SRSV strain. Comparison of the RNA polymerase region of this virus with the equivalent regions of genetic group I (69.4‐75.0% amino acid identity) and genetic group II (88.9‐100% amino acid and 77.1‐88.1% nucleotide identity) SRSVs revealed that the causative SRSV was a distinct member of genetic group II. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus genotypes in belgian hemophiliacs
Authors: Sheng Li , Willems Marc , Peerlinck Kathelijne , Yap Sing Hiem , Vermylen Jozef
Published: March 2007

Hemophiliacs who have been exposed to un‐heated and/or dry heated pooled clotting factor concentrates are at a high risk of chronic hepatitis C. Serum HCV‐RNA was measured by reverse transcriptase‐polymerase chain reaction (RT‐PCR) technique in 58 hemophiliacs positive for anti‐HCV antibodies, HCV‐RNA was detected in 55 patients. Based on the results of PCR positiv‐ity, HCV genotyping was carried out using genotype specific probes and the dot blot hybridization assay: 2 patients were found to be infected with type 1a (3.6%), 31 patients with type 1b (56.4%), 8 patients with type 2 (14.6%), 6 patients with type 3 (10.9%), 1 patient with type 4, and 7 patients with double infection (12.7%). As compared to a control group of 105 nonhemophilia patients with chronic hepatitis C, HCV genotype 1 b was found predominant in both groups. However, double infection with two HCV types as well as a relatively high prevalence of infection with type 2 and type 3 were found in hemophiliacs. © 1995 Wiley‐Liss, Inc.
Molecular characterization of a human calicivirus with sequence relationships closer to animal caliciviruses than other known human caliciviruses
Authors: Matson David O. , Zhong Wei‐Ming , Jiang Xi , Pickering Larry K. , Nakata Shuji , Numata K. , Chiba Shunzo , Estes Mary K.
Published: March 2007

cDNA clones were produced from a morphologically typical human calicivirus (HuCV) in stool specimens collected in 1982 during an outbreak of gastroenteritis in Sapporo, Japan. The cDNA clones were generated separately in two laboratories by reverse transcriptase‐polymerase chain reaction (RT‐PCR) using primers 35 and 36 derived from Norwalk virus. The RT‐PCR product from six specimens was of the predicted size, had a continuous protein encoding frame on the positive strand, and contained GLPS and YGDD amino acid motifs at the predicted distance from the primers. RT‐PCR amplification with primer 35 and a HuCV/Sapporo‐specific primer 36 of four HuCV/Sapporo‐positive stool specimens from a 1986 Houston day care center outbreak yielded products with 93% nucleotide and 99% predicted amino acid sequence identity with the HuCV/ Sapporo strain from the 1982 outbreak. The HuCV/Sapporo strains are genetically distinct from previously characterized HuCVs and more closely related to known animal CVs than other known HuCVs. © 1995 Wiley‐Liss, Inc.
Detection of hepatitis C virus RNA in the cell fraction of saliva before and after oral surgery
Authors: Chen Margaret , Yun Zhi‐Bing , Sonnerborg Anders , Schvarcz Robert , Bergquist Inge , Berghmd Hans‐Bertil , Sallberg Matti
Published: March 2007

The presence of hepatitis C virus (HCV) RNA in serum, whole saliva, and saliva from the sub‐maxillary glands was investigated before and after oral surgery‐ The presence of HCV RNA (positive and negative‐strand RNA) was determined in serum and saliva by a nested polymerase chain reaction in 26 anti‐HCV positive patients, of whom 11 were coinfected with human immunodeficiency virus‐1. Oral surgery was carried out on five occasions on four of the patients. HCV RNA was detected in the sera of 23 of 26 (88%) patients, and in the saliva of 4 of the 23 (17%) of the viremic patients. In all four cases, HCV RNA was detected only in the cell fraction derived from centrifugation of whole saliva. Negativestranded HCV RNA was not detected. At one of five occasions of oral surgery, HCV RNA was detected in saliva sampled immediately after surgery, but not before or 24 hours after surgery. The results suggest that HCV is present in saliva in less than 25% of HCV viremic persons. The presence of the virus in saliva is restricted to the cell fraction. Thus, saliva may serve as a possible, but low, nonparenteral transmission route of HCV. Contamination of saliva by blood during and after oral surgery may result in an increased risk of viral exposure. Except for trauma caused by sharp instruments during surgery, this might contribute to the higher HCV seropositivity found among dentists. © 1995 Wiley‐Liss, Inc.
Measles antibodies in women and infants in the vaccine era.
Authors: Kacica Marilyn A. , Hughes Patricia A. , Lepow Martha L. , Venzia Richard A. , Miller Joan
Published: March 2007

The present investigation was done to determine whether measles enzyme immune assay (EIA) absorbency values were lower in women born in the vaccine era after 1963 and their infants in an upstate New York metropolitan area, an area of low measles incidence during the past 10 years compared with women born before the measles vaccine era who had natural measles. Aliquots of 202 sera from mother‐infant pairs collected for other purposes from November 1990 to June 1991 at Albany Medical Center Hospital were tested by EIA. The demographic data available for analysis were maternal age and infant gestational age. Measles mean absorbency values were analyzed according to maternal age. Of 202 mother‐infant pairs, 30% of mothers and 17% of their infants were seronegative (EIA < 0.16). Mothers born before 1963 and their infants had significantly higher mean EIA absorbency values than mothers born after 1963 and their infants (P < 0,002). The percent seropositive for measles antibodies by EIA for mothers born before 1963 and their infants, 87% and 94%, respectively, was significantly higher than the percent seropositive for mothers born after 1963 and their infants, 61% and 69%, respectively (P = 0.0001). Since the mean measles antibodies as measured by EIA absorbency were significantly lower in the mothers born after 1963 and their infants compared with women born before the vaccine era, the strategy for measles control in the future may have to include lowering the age of infant immunization. © 1995 Wiley‐Liss, Inc.
Fulminant human herpesvirus six encephalitis in a human immunodeficiency virus‐infected infant
Authors: Knox Konstance K. , Harrington Daniel P. , Carrigan Donald R.
Published: March 2007

Self‐limited involvement of the central nervous system (CNS) is a relatively common complication of primary infection with human herpesvirus six (HHV‐6) in normal children. We describe an HIV‐infected infant who developed fulminant encephalitis as a complication of HHV‐6 infection. Immunohistochemical staining of CNS tissue demonstrated productive infection of all CNS cell‐types. Analysis of the infected brain tissue by the polymerase chain reaction (PCR) confirmed the presence of a dense HHV‐6 infection in the tissue, and demonstrated that the virus present in the CNS tissue was predominantly the A variant of HHV‐6. This is the first demonstration of invasive tissue disease caused by HHV‐6 in an HIV‐infected infant. © 1995 Wiley‐Liss, Inc.
Screening a monoclonal antibody with a fusion‐phage display library shows a discontinuity in a linear epitope within pres1 of hepatitis B virus
Authors: Germaschewski Volker , Murray Kenneth
Published: March 2007

The epitope recognized by the monoclonal antibody MA18/7, specific for the PreS1‐domain of the hepatitis B virus surface antigen, has been defined precisely by means of a library of fusionphage carrying random hexapeptides on the tip of filamentous phage fd particles. Phage, isolated after only one round of affinity selection, displayed hexapeptides showing strong conservation of the PreSl primary sequence in the region 19–23 with three noncontiguous residues, DP (20 and 21) and F (23) appearing in phage that bound the antibody. The importance of these core residues was supported by comparing the antibody binding of individual phage in solution, which provided relative dissociation constants for these interactions. Replacement of F (23) by Y was the only substitution observed in the three core residues, and resulted in somewhat weaker binding. Synthetic tetra‐ and hexapeptides containing these key residues inhibited the reaction between the phage and the antibody. © 1995 Wiley‐Liss, Inc.
Serological evidence for hepatitis E virus infection in Israel
Authors: Karetnyi Yuory V. , Handsher Rachel , Aboudy Yair , Varsano Noemi , Mendelson Ella , Weiss Peretz , Levin Edit , Bar‐Shani Shulamit , Schwartz Eli , Fields Howard A. , Favorov Michael O. , Khudyakova Natasha S.
Published: March 2007

Israel is suspected to be endemic for hepatitis E virus (HEV) because of its geographic location and the large‐scale immigration from endemic countries. Although no cases of local HEV infection have been diagnosed, a serological survey would provide indirect evidence for such infection. We examined sera from 1,416 healthy subjects, including 1,139 Jews from various regions of Israel and 277 Arabs, most of whom reside in the West Bank of the Jordan River. In addition, we tested 13 non‐A, non‐B, and non‐C viral hepatitis patients. Sera were screened for antibody to hepatitis E virus (anti‐HEV) by a newly developed enzyme immunoassay (EIA) and by immuno‐blots for both IgG and IgM anti‐HEV activity. Positive samples were confirmed by neutralization.

The seroprevalence found by EIA was 2.81% and 1.81% in the Jewish and Arab populations, respectively. More than a 2‐fold higher prevalence in males compared to females and an increase with age were found in both populations. However, these differences were nonsignificant. The geographical distribution was even throughout the country, except for two clusters of 3 and 4 seropositive individuals possibly reflecting past foci of infection. Eight of 37 ElA‐positive sera were positive for IgG, and 3 were positive for IgM by the immunoblot assay. Among hepatitis patients (9 acute and 4 chronic), one patient with chronic hepatitis was positive for both IgG and IgM.

Our study provides indirect evidence that Israel is endemic for HEV. The lack of outbreaks may be attributed to generally good hygienic conditions and a controlled potable water supply, while unrecognized sporadic cases may be due to the unavailability of diagnostic tests. © 1995 Wiley‐Liss, Inc.
Genetic heterogeneity in the precore region of hepatitis B virus in hepatitis B e antigen‐negative chronic hepatitis B Patients: Spontaneous seroconversion and interferon‐induced seroconversion
Authors: Karasawa Tatsunobu , Aizawa Yoshio , Zeniya Mikio , Kuramoto Akira , Toda Gotaro , Shirasawa Takuji
Published: March 2007

To elucidate the relationship between the clinical severity of chronic liver disease and the precore mutations in hepatitis B e antigen (HBeAg)‐nega‐tive hepatitis B virus (HBV) carriers, mutations were investigated in the precore region of HBV DNA in 20 chronic hepatitis B patients who sero‐converted either spontaneously or after the administration of α‐interferon (IFN), and 5 asymptomatic carriers. The precore mutation with a stop codon at nucleotide 1896 was found in all patients, irrespective of the histology and in all asymptomatic carriers. The second mutation at nucleotide 1899 was found in 40% of cases studied but always followed by the first mutation at nucleotide 1896. The mixed viral infection of precore mutant and wild‐type HBV virus was found in 40% of seroconverted cases after IFN treatment and in sera of HBV carriers obtained within a year after the spontaneous Seroconversion. These data suggest that the precore mutants prevail over wild‐type HBV in all HBeAg‐negative HBV carriers within several years after the sero‐conversion, but their prevalence could not confine the clinical severity of chronic liver disease. © 1995 Wiley‐Liss, Inc.
Presence of human T‐lymphotropic virus type II‐related genes in DNA of peripheral leukocytes from patients with autoimmune thyroid diseases
Authors: Yokoi Kenji , Kawai Hisaomi , Akaike Masashi , Mine Hideki , Saito Shiro
Published: March 2007

No etiologic role of human T‐lymphotropic virus type II (HTLV‐II) in any disease is yet known. The present study showed a high incidence of HTLV‐II proviral DNA fragments in DNA of peripheral blood leukocytes from patients with autoimmune thyroid diseases. Using primers for the pol and tax regions of HTLV‐II proviral DNA, amplified DNA fragments were demonstrated in 51.5% of the patients with Hashimoto's thyroiditis and in 11.8% of those with Graves' disease examined, but in only 1.9% of the disease controls and 1.0% of healthy individuals. These amplified DNA fragments hybridized with each of the inner probes. The nucleotide sequences of the DNA fragments of the pol and tax regions showed high homology with those of the prototype of HTLV‐II. No antibodies for HTLV‐II could, however, be detected in the patients examined. Because of the absence of its antibody, HTLV‐II infection was not confirmed in these patients, but the presence of HTLV‐II proviral DNA or its related DNA at high frequency suggests a relationship of HTLV‐II with the development of autoimmune thyroid diseases. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus genotypes in French blood donors
Authors: Gournay Jérǒme , Marcellin Patrick , Martinot‐Peignoux Michèle , Gabriel Franck , Erlinger Serge , Benhamou Jean‐Pierre , Degott Claude , Courtois Françoise , Branger Michel , Wild Anne Marie
Published: March 2007

The prevalence of different hepatitis C virus (HCV) genotypes in HCV infected individuals and the relation between the HCV genotypes and the source of the infection are controversial. The aim of this study was to determine the HCV genotypes in French blood donors. Fifty‐one anti‐HCV positive blood donors were studied with detectable serum HCV RNA by nested polymerase chain reaction (PCR) using primers derived from the 5′ non‐coding region. For genotyping HCV, we used a method based on analysis of the restriction fragment length polymorphisms (RFLP) after amplification by PCR of the HCV non‐structural 5 (NS5) genome domain. Using this technique, the genotypes of 39 of the 51 blood donors (76%) were determined. Three previously described genotypes were found: 19 blood donors were infected by HCV genotype I (37%), 14 were infected by HCV genotype II (27%), 3 were infected by HCV genotype III (6%), and 3 were coinfected by two genotypes (6%). All three blood donors infected with two different genotypes were intravenous drug abusers. A past history of intravenous drug abuse was more frequent in blood donors with HCV genotype I. However, there was no difference in alanine transaminase (ALT) levels, histological lesions, and RIBA‐2 patterns in blood donors infected with either HCV genotype I or HCV genotype II. These findings indicate that most HCV genotypes isolated from French blood donors belong to HCV genotype I and HCV genotype II, and that risk factors for HCV infection may differ for different genotypes of HCV. © 1995 Wiley‐Liss, inc.
Interferon therapy in chronic hepatitis C virus: Evidence of different outcome with respect to different viral strains
Authors: Pozzato Gabriele , Moretti Michèle , Crocé Lory Saveria , Sasso Franco , Tiribelli Claudio , Crovatto Marina , Santini Gianfranco , Kaneko Suichi , Unoura Masashi , Kobayashi Kenichi
Published: March 2007

The aim of the study was to assess the role of different viral strains of hepatitis C virus (HCV) in determining the outcome of the alpha‐interferon (IFN) therapy. Fifty‐seven patients (34 from Italy and 23 from Japan) with HCV‐positive liver disease were enrolled in the study. The NS4 region of HCV was amplified in sera by “nested” polymerase chain reaction (PCR) using a primer pair synthesized according to the sequence of JK‐1. The NS4 region was positive in 14 (41%) Italian and in 13 (56%) Japanese patients. In positive patients the sequence of the NS4 region was also obtained. Subsequently, HCV genotype was determined in all patients by PCR amplification of the core region. All patients received recombinant alpha2a‐interferon (IFN), 6 million units 3 times a week for 1 month followed by 3 million units 3 times a week for 5 months. The patients were followed for 1 year after the end of treatment. At the end of the follow‐up, 17 (30%) had sustained normal levels of serum alanine aminotransferase (ALT). The outcome of treatment was not correlated with race, age, sex, histology, and pretreatment ALT level, but was significantly (P < 0.00001) associated with the presence of both the NS4‐JK‐1 region and HCV type II. Among the 27 NS4‐positive patients, only 1 patient (3.7%) achieved a complete response, whereas the remaining 26 patients (96.3%) either were non‐responders or relapsed after IFN was discontinued. In contrast, among the 30 NS4‐JK‐1‐negative patients, 15 (53%) had a sustained remission. HCV genotyping showed type I in 3 (6%), type II in 40 (74%), type III in 4 (7%), and type IV in 3 (6%) cases. Coinfection was present in 4 (7%), while in 3 cases amplification was not obtained. Patients with type II were all non‐responders or relapsers, while a response to the treatment was oberved in 17 of 17 (100%) of the remaining patients. These data indicate that the presence of JK‐1 variant of HCV or HCV type II is almost always predictive of a poor response rate Of IFN therapy. © 1995 Wiley‐Liss, Inc.
Antibodies that block virus attachment to vero cells are a major component of the human neutralizing antibody response against dengue virus type 2
Authors: He Run‐Tao , Wang Songli , Anderson Robert , Innis Bruce L. , Nisalak Ananda , Usawattanakul Wipawee , And Siripen Kalayanarooj
Published: March 2007

Epidemiological data strongly implicate a role for the host humoral immune response in both protection against and exacerbation of dengue virus‐caused disease. In an effort to characterize elements of the normal human immune response against dengue virus we have addressed the issue of antibody‐mediated neutralization of dengue virus. We show here the ability of both mouse monoclonal antibody 3H5 and human anti‐dengue neutralizing sera to block binding of dengue‐2 virus to monkey kidney (Vero) cells. Since Vero cells possess virus receptors but not Fc receptors we conclude that the major effect of host neutralizing antibodies is to block virus attachment to Vero cell dengue virus receptors. Analysis of 61 patient antisera yielded good correlation (Pearson's coefficient = 0.90; P < 0.001) between neutralizing activity and ability to block virus‐cell attachment suggesting that antibody‐mediated neutralization of dengue virus occurs primarily extracellularly and less by a postat‐tachment mechanism as has been described for certain other viruses. © 1995 Wiley‐Liss, inc.
Mapping of a human T‐lymphotropic virus type I gag protein epitope that cross‐reacts with anti‐Plasmodium falciparum antibodies
Authors: Porter Kevin R. , Hayes Curtis G. , Anthony Ronald L. , Solihin Ating
Published: March 2007

Studies in Palawan, Philippines, and Irian Jaya, Indonesia, showed that indeterminate human T‐lymphotropic virus type I (HTLV‐I) Western blot immunoreactivity is due to cross‐reacting anti‐Plasmodium falciparum antibodies. To further define this immunoreactivity, mapping studies were conducted using the HTLV‐I p19 protein to identify the precise epitope that reacts with these antibodies. Anti‐P. falciparum antibody‐positive sera from Palawan, Philippines, and Irian Jaya, Indonesia, were studied using overlapping synthetic peptides. Immunoreactivity was localized to residues 108–120 of p19. Further analysis of the sera with 5 biotinylated synthetic peptides showed that the cross‐reactive epitope consists of the sequence PDSDPQI (amino acid residues 110–116), which was shown to be homologous to a 7 amino acid sequence on the Exp‐1 protein of the P. falciparum blood stage parasite. This is the first study that identifies a specific HTLV‐I protein epitope that cross‐reacts with malaria antibodies. © 1995 Wiley‐Liss, Inc.

This article is a US Government work and, as such, is in the public domain in the United States of America.
Correlation of human papillomavirus 16 and 18 with prognostic factors in invasive cervical neoplasias
Authors: Kónya József , Veress György , Hernádi Zoltán , Soós Györgyike , Czeglédy Judit , Gergely Lajos
Published: March 2007

Forty‐seven patients with cervical carcinoma were examined in order to correlate human papillomavirus (HPV) types with prognostic factors in invasive cervical neoplasias. Age, clinical stage, histological type, and grade and parity were analysed with respect to HPV status as determined by a general primer mediated polymerase chain reaction (PCR) or a type specific PCR. All but one sample (98%) harboured HPV sequences: HPV 16 was found in 26 cases (55%), HPV 18 in 19 cases (40%0), and HPV 31 in 1 case. The presence of HPV 18 DNA was significantly associated with cancers developed below 40 years of age (P = 0.029). HPV 18 detection was associated with poor differentiation malignancy (P = 0.045) and histological types of poor prognosis (adenocarcinoma or nondifferentiated carcinoma; P = 0.006). HPV 18 positivity was also correlated with advanced clinical stages (FIG0 II and III; P = 0.032). Parity and HPV status proved to be independent of each other (P – 0.99). Eighty‐seven percent (27/31) of pelvic lymph nodes from HPV positive patients contained HPV DNA. The virus types found in lymph nodes were identical with those of the primary tumours in all cases. Virological results were compared to those obtained by routine histological examination. Only 6 of 27 patients with HPV positive lymph nodes had any histological evidence of metastasis. Nevertheless, the lack of metastasis as detected by histology does not exclude the possibility of relapses. Follow‐up of the clinical course of the disease is required to assess the prognostic significance of PCR detection of HPV in the possible sites of early metastases. © 1995 Wiley‐Liss, Inc.
Circulating immune complexes in chronic hepatitis C
Authors: Tsai Jung‐Fa , Jeng Jen‐Eing , Chang Wen‐Yu , Ho Mei‐Shang , Lin Zu‐Yau , Tsai Juei‐Hsiung
Published: March 2007

For assessing the role of circulating immune complexes (CIC) in chronic hepatitis C, the relative frequency of CIC was determined in 54 patients with chronic hepatitis C, 15 asymptomatic hepatitis C virus (HCV) carriers, and 54 healthy controls. IgM and IgG containing CIC were studied using both Clq and conglutinin (K) in an immunoglobulin‐specific solid‐phase enzyme immunoassay. CIC were a common feature of chronic hepatitis C with 96.3% of patients with at least one abnormal test result. The prevalence of elevated IgG‐K, IgM‐K, IgG‐C1q, and IgM‐C1q CIC was 70.3%, 50.0%, 64.8%, and 35.1%, respectively. The prevalence of IgG class CIC was higher than IgM class CIC (P = 0.038 for K‐CIC and P = 0.01 for C1q‐CIC, respectively). There is correlation between IgG‐K CIC and IgG‐C1q CIC (r = 0.445, P= 0.0021, IgG‐K CIC and ISM‐Clq CIC (r = 0.348, P = 0.020). IgM‐K CIC and aspartic arninotransferase (r = 0.321, P= 0.015). IgM‐K CIC and alanine aminotransferase (r =0.301, P = 0.027). Compared to patients with chronic persistent hepatitis and chronic lobular hepatitis, patients with chronic active hepatitis have a higher prevalence of elevated IgG‐K CIC (77.2% vs. 40.0%, P = 0.029) and IgM‐K CIC (56.8% vs. 20.0%, P = 0.038). The concentration of IgG‐K, IgM‐K, and IgM‐C1q CIC in the former was significantly higher than that in the latter, respectively. In conclusion, IgG class CIC is the major type of CIC in chronic hepatitis C. Conglutinin‐binding CIC correlates with more severe tissue damage. CIC may play a role in the pathogenesis of chronic hepatitis C. © 1995 Wiley‐Liss, Inc.
Phylogenetically distinct hantavirus implicated in a case of hantavirus pulmonary syndrome in the Northeastern United States
Authors: Hjelle Brian , Krolikowski John , Torrez‐Martinez Norah , Chavez‐Giles Francesca , Vanner Cynthia , Laposata Elizabeth
Published: March 2007

Hantavirus pulmonary syndrome (HPS) is an acute respiratory illness with high mortality. It is caused by a newly described New World hantavirus known as Four Corners virus (FCV). Nearly all cases of HPS have occurred in the western United States. The etiologic agents in those cases have been closely related to each other, based upon comparisons of their genetic sequences. We have molecularly cloned the S genomic segment of a hantavirus (RI‐1) implicated in a case on HPS in the northeastern United States. Nucleotide sequence analysis shows that the RI‐1 virus has many similarities to FCV, but is clearly distinct from the western forms of that virus. These data suggest that HPS can be caused by multiple agents that together form a distinctive evolutionary cIade. © 1995 Wiley‐Liss, Inc.
Isolation of black creek canal virus, a new hantavirus from Sigmodon hispidus in Florida
Authors: Rollin Pierre E. , Ksiazek Thomas G. , Elliott Luanne H. , Ravkov Eugene V. , Martin Mary Lane , Morzunov Sergey , Livingstone Walter , Monroe Marty , Glass Gregory , Ruo Sup , Khan Ali S. , Childs James E. , Nichol Stuart T. , Peters C. J.
Published: March 2007

Numerous rodents were trapped for serologic and virologic studies following the identification of a hantavirus pulmonary syndrome (HPS) case in Dade County, Florida. Cotton rats (Sigmodon hispidus) were the most frequently captured rodent and displayed the highest seroprevalence to a variety of hantavirus antigens. Hantavirus genome RNA was detected in all the seropositive cotton rats tested, using a reverse transcriptase‐polymerase chain reaction (RT‐PCR) assay. A virus was isolated from tissues of two seropositive cotton rats by cultivation of lung and spleen homogenates on Vero E6 cells. Nucleotide sequence information obtained by direct RT‐PCR and the serologic relationships of this virus with the other hantaviruses indicate that this virus, Black Creek Canal virus, represents a new hantavirus distinct from the previously known serotypes. © 1995 Wiley‐Liss, Inc.
Combined treatment with interferon alpha‐2b and ribavirin for chronic hepatitis C in patients with a previous non‐response or non‐sustained response to interferon alone
Authors: Schvarcz R. , Yun Z. B. , Sönnerborg A. , Weiland O.
Published: March 2007

Ten patients with chronic hepatitis C, six of whom had not responded and four of whom had responded in a non‐sustained fashion to interferon‐alpha treatment alone, were given interferon alpha‐2b and ribavirin in combination during 24 weeks. Interferon alpha‐2b was given subcutaneously, at a dose of 3 MU thrice weekly, together with ribavirin orally, at a dose of 1, 000‐1, 200 mg/day.

All four patients with a prior non‐sustained response to interferon alone had normal alanine aminotransferase (ALT) levels at the end of treatment as well as during follow‐up (⩾24 weeks post treatment). Furthermore, all four lost serum HCV‐RNA at the end of treatment and three continued to be negative during follow‐up. Among patients with a prior non‐response to interferon alone three of six had normal ALT levels at the end of treatment and one at follow‐up. Two of six became HCV‐RNA negative at cessation of treatment, one of whom was negative also at followup. All former non‐sustained responders and one of six non‐responder patients thus showed a sustained biochemical response with eradication of HCV‐RNA from serum in all cases but one.

It is concluded that combination therapy with interferon alpha‐2b and ribavirin offers a chance of sustained biochemical resposne with eradication of the viremia in patients who have not shown a persistent response to interferon‐alpha alone. © 1995 Wiley‐Liss, Inc.
Detection of human herpesvirus 6 DNAs in samples from several body sites of patients with exanthem subitum and their mothers by polymerase chain reaction assay
Authors: Suga Sadao , Yazaki Takehiko , Kajita Yqji , Ozaki Takao , Asano Yoshizo
Published: March 2007

Polymerase chain reaction amplification was used to detect human herpesvirus 6 (HHV‐6) DNAs in peripheral blood mononuclear cells (MNCs), plasma, saliva, stool, and urine from three patients with exanthem subitum and in peripheral blood MNCs, plasma, and saliva from their mothers. HHV‐6 DNAs were detected in MNCs during and after the disease and were found in plasma only in the acute phase. The virus DNAs were also detected in saliva after recovery from the illness and were found persistently or intermittently in stool but not in urine samples after the onset of the disease. In contrast, one of the three mothers excreted HHV‐6 DNAs persistently in saliva. None of the mothers had the virus DNAs in peripheral blood MNCs and plasma nor a significant increase in antibody titers to HHV‐6 after possible exposure from their children. These findings suggest systemic replication of HHV‐6 during the acute phase in patients with exanthem subitum and persistent infection of the virus in several organs after recovery from the disease. © 1995 Wiley‐Liss, Inc.
Persistent B19 parvovirus infections in hemophilic HIV‐1 infected patients
Authors: Musiani M. , Zerbini M. , Gentilomi G. , Rodorigo G. , De Rosa V. , Gibellini D. , Venturoli S. , Gallinella G.
Published: March 2007

B19 infection can be acquired by transmission with blood factors in patients with congenital bleeding disorders, requiring clotting factor concentrates. In immunodeficient patients, the failure of immunity to clear B19 virus may produce persistent infections. The presence of B19 DNA in blood samples from seven hemophilic patients with concomitant HIV‐1 infection was studied over a period of three‐to‐four years. Dot blot hybridization assays with DNA and RNA probes were used to detect medium high viremias, and polymerase chain reaction (PCR) to detect very low viremic titres. Three patients were negative for B19 DNA in all the blood samples, while four patients were persistently positive for B19 DNA. Viral persistence, which in one patient was detected throughout the study period (40 months), occurred at low titre in all four positive patients with some recurrent increases in viral titre. In the four patients persistently positive for B19 DNA, acute or chronic clinical symptoms and signs that could be associated with B19were not noted when virus was present at low titre (B19 DNA detectable only by PCR). When patients had a higher viral titre (B19 DNA detectable by dot blot hybridization) acute manifestations (aplastic crisis, Fifth disease, fevers, pneumonitis) were found. © 1995 Wiiey‐Liss, Inc. © 1995 Wiley‐Liss, Inc.
Peripheral lymphocytes of clinically non‐progressor patients harbor inactive and uninducible HIV proviruses
Authors: Garbuglia A. Rosa , Salvi Roberto , Caro Antonino Di , Montella Francesco , Di Sora Fiorella , Recchia Olga , Delfini Carlo , Benedetto Arrigo
Published: March 2007

The HIV viral burden and RNA expression in a selected group of infected, clinically non‐progressor patients were investigated. Five fast‐progressor patients and 10 AIDS cases were included as controls. The HIV viral load was investigated by semiquantitative polymerase chain reaction (PCR) in adherent macrophages and in genomic and extragenomic fractions of lymphocytes. HIV DNA was not found in macrophages in the non‐progressor subjects, was weakly positive in 2 of 5 fast‐progressors and strongly positive in most of the AIDS patients. The number of HIV proviruses found in lymphocytes of the non‐progressor subjects varied from 5 to 160 copies/μg DNA, values ten times lower than those recorded in fast‐progressors and AIDS patients. The extragenomic HIV DNA (2 LTR forms) was absent or barely detectable in the lymphocytes from non‐progressors and abundant in the other groups. HIV RNA was not found in the lymphocytes of all non‐progressors. This may indicate that a latent state of HIV provirus exists in the lymphocytes of these subjects. To investigate this point, cultivation and stimulation with PHA (phytohemoagglutinin) and PMA (phorbol 12‐myristate 13‐acetate) of lymphocytes from these subjects were attempted but after 6 days HIV RNA (RT‐PCR for gag region) was still absent or barely detectable in these patients. There are no other reports of the absence of HIV provirus induction in lymphocytes from infected individuals. If confirmed in a larger number of patients, such non‐inducibility might serve as a predictor marker of progression of the disease. © 1995 Wiley‐Liss, Inc. © 1995 Wiley‐Liss, Inc.
Polymerase chain reaction for rapid detection of ocular adenovirus infection
Authors: Morris D. J. , Bailey A. S. , Cooper R. J. , Turner P. C. , Jackson R. , Corbitt G. , Tullo A. B.
Published: March 2007

Adenoviruses are associated with endemic and epidemic acute conjunctivitis, large nosocomial outbreaks reflecting virus transmission on unwashed hands or inadequately sterilised ophthalmic instruments. The polymerase chain reaction (PCR) proved more sensitive than antigen detection by immune dot‐blot test for the rapid diagnosis of ocular adenovirus infection (sensitivities in a retrospective study 112/123 (91%) versus 72/123 (59%), P< 0.001). Indeed, in a prospective comparison, DNA amplification and virus isolation generated similar numbers of positive results (34 versus 32), though five PCR positive results were possibly false positives. The sensitivity of the PCR was largely independent of adenovirus subgenus or serotype, though reduced sensitivity with subgenus B strains could not be excluded. Specimen preparation for DNA amplification using a simple lysis buffer proved more effective than phenol‐chloroform extraction. The immune dot‐blot test gave unavoidable false positive results, but with the PCR this problem could be minimized by technical modifications. The PCR could replace antigen detection and virus isolation as the initial test for adenoviruses in conjunctival swabs, with cell culture only being retained for adenovirus serotyping in PCR positive specimens and for other viruses such as herpes simplex. © 1995 Wiley‐Liss, Inc.
Varicella‐zoster virus assembly protein p32/p36 is present in DNA‐containing as well as immature capsids
Authors: Harper D. R. , Sanders E. A. , Ashcroft M. A.
Published: March 2007

Varicella‐zoster virus (VZV) produces a group of nucleocapsid proteins (the p32/p36 nucleoprotein complex) which are the VZV analogues of the herpes simplex virus type 1 (HSV‐1) and cy0tomegalovirus (CMV) assembly proteins. There are multiple components in the VZV p32/p36 complex, with major proteins of 32 and 36 kDa and minor proteins of 34 and 38 kDa. In HSV‐1 the assembly proteins have been shown to be present in immature (B) capsids, but are removed prior to the formation of mature (C) capsids containing the viral DNA genome. Our work has shown that VZV produces capsids corresponding to the B and C forms. However, in contrast to HSV‐1, VZV also produces “B/C” capsids that appear to contain both the assembly proteins and the viral DNA genome. Possible mechanisms for this are discussed.

In addition, it was shown that VZV capsids appear to lack the 36 and 38 kDa proteins, and based on this observation we suggest that these may represent unprocessed forms of the assembly protein.

In both HSV and CMV, a much larger, crossreactive protein has been identified as the fulllength product of the gene coding for the assembly protein. The homologous VZV gene (ORF 33) theoretically has the capacity to produce a 66 kDa protein. However, no such protein is readily apparent in VZV‐infected cells. The presence of an immunoreactive 64 kDa protein was demonstrated in purified VZV capsids which may represent the full‐length ORF 33 protein. © 1995 Wiley‐Liss, Inc.