bluuebunny/crossref_metadata_2025 huggingface dataset:
An enzyme‐linked immunosorbent assay (ELISA) for IgG and IgA antibodies to respiratory syncytial virus in low dilutions of secretions of human serum and secretions
Authors: Nandapalan N. , Routledge E. , Toms G. L.
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) has been developed for titration of IgG and IgA antibodies to respiratory syncytial (RS) virus in low dilutions of human serum, colostrum, and nasopharyngeal secretions. Previously the sensitivity of RS virus ELISA on such specimens has been limited by nonspecific absorption of antibody, particularly IgA, to crude antigen preparations.

For IgG antibody estimation in infant sera, this unwanted binding was reduced to workable levels by increasing the serum, salt, and detergent concentration of the diluent. Residual nonspecific binding of IgA in colostra appeared mainly due to antigen lipids or to lipoproteins. This was markedly reduced by partitioning Triton X‐100‐treated infected cell lysate antigens in Arklone.

Using the modified ELISA technique for anti‐RS virus IgA, good correlations were found with unfixed cell membrane immunofluorescence (MIF) for colostra (r = 0.81, P < 0.001) and nasal secretions from adult volunteers. In several samples nonspecific absorption of antibody precluded MIF assay, but did not affect the ELISA. Although there was an overall correlation between ELISA for anti‐RS IgG antibody in sera, the complement fixation test (r = 0.75, P < 0.001), and MIF test (r = 0.82, P < 0.001), the sensitivity of ELISA for antibody responses in convalescent sera of infants from 3 months to 2 years was poor. Conversely, the sensitivity of ELISA for antibody in the sera of older children and for transplacentally acquired antibody in very young infants was higher than that for the other two tests. ELISA was thus less reliable than either CF or MIF for detecting antibody rises in paired infant sera, particularly where maternally acquired antibody remained in the acute serum. The reasons for this apparent disparity are discussed.
Cellular immune response to infection with respiratory syncytial virus and influence of breast‐feeding on the response
Authors: Mito Kazuaki , Chiba Yasuo , Suga Kazuhiro , Nakao Tooru
Published: March 2007

Virus‐specific lymphocyte transformation (LTF) activity in vitro was examined in 78 infants with various forms of illness due to respiratory syncytial virus (RSV) infection. In 73 subjects with lower respiratory tract disease, significant LTF activity was often detected within one week after onset of initial symptoms, and responses characteristic for each clinical form were observed in the subsequent rise. Thus, mean activity in subjects with tracheobronchitis increased gradually, with the maximum response being detected at the fourth week after the onset of illness. In subjects with pneumonia the response was rather low during the first week of illness, and then a sharp increase of activity was observed at the second week. Although patients with bronchiolitis elicited similar levels of the activity until the first week, the response was significantly suppressed during the subsequent two weeks. The response at the second week in this group was significantly lower than those of corresponding specimens obtained from patients with pneumonia (p < 0.01) or tracheobronchitis (p < 0.05), suggesting a close association of the responsiveness with underlying mechanisms of bronchiolitis induced by RSV. The present study further indicated a possibility that breast‐feeding of RSV‐infected infants may alter levels of the LTF activity in these subjects.
Local and peripheral cell‐mediated immune response to influenza virus in mice
Authors: Shapira‐Nahor Orit , Zakay‐Rones Zichria
Published: March 2007

Presentation of different influenza virus antigens generates different immune responses. Intranasal immunization with either live (VA) or formalin‐inactivated (VF) A/PR/8/34 (HoN1) influenza virus induced local as well as peripheral cell‐mediated immune response (CMI), as evidenced by elevation in 3H‐thymidine incorporation. Cell‐mediated immune response was detected as soon as 24–48 hr following the application of VA and 4–5 days following VF. Cell‐mediated immune response in both instances peaked on the 12th day and disappeared between 16 and 20 days after application.

Local CMI response was threefold higher after immunization with VA (SI = 28.6) than with VF (SI = 9.4), while VF induced higher peripheral response (32.0 vs 17.7). The mononuclear cell population in the lungs increased, correlating with a rise in the stimulation index (SI). The percentage of IgA surface‐bearing B lymphocytes was significantly higher following IN administration of VA, but not following VF instillation. This corroborated the finding that VF failed to induce local antibody response in the lungs in spite of its capacity to stimulate humoral antibody and CMI responses.

Mice immunized intramuscularly with both viral preparations developed a fair humoral antibody response without detectable CMI (peripheral or local).
Attenuated Junin virus infection in callithrix jacchus
Authors: Avila Dra. M. M. , Frigerio M. J. , Weber E. L. , Rondinone S. , Samoilovich S. R. , Laguens R. P. , de Guerrero L. B. , Weissenbacher M. C.
Published: March 2007

Twenty marmosets, male Callithrix jacchus, were used during this study. Fifteen of the marmosets were inoculated with 5,000 TCID50 of the attenuated XJCl3 strain of Junin virus by intramuscular route and five were left as uninoculated controls. Animals were observed for a 420‐day period. In order to carry out virologic, hematologic, serologic, and histologic studies the animals were bled and/or killed at different days post infection(pi).

Results obtained showed that the attenuated strain produced an infection with no mortality or signs of illness. There was only a slight loss of weight at 18–40 days pi, which was soon recovered. Viremia was present from day 6 to 22, titers peaking at 4.0 log. Viral spread was limited to the lungs, spleen, lymph nodes, and bone marrow in the animal killed on day 14. No virus was found in the organs of the animal killed on day 23, and neither hematologic alterations nor pathologic lesions were seen in these monkeys except for ganglionar hypertrophy with immunoblast proliferation. Antigen was detected by immunofluorescence (IF) in lymph nodes, spleen, adrenals, lungs and brain.

Neutralizing antibodies were detected from the third week onward. Protection conferred by the XJC13 strain proved effective when XJCU–inoculated monkeys were challenged with 1,000 TCID50 of the pathogenic XJ strain at days 60 or 380 pi, while normal controls died.

When viral persistence was searched for on days 370, 390, and 420 pi, no infectious virus was detected, but viral antigen was seen in certain organs, which, however, lacked tissue damage.

Bearing in mind the main features differentiating the previously tested pathogenic and the attenuated strains, C jacchus may prove a useful model for testing the pathogenicity and efficacy of attenuated vaccines against Argentine hemorrhagic fever (AHF).
Neurotropism of a high‐passage XJ strain of junín virus
Authors: Oubiña José R. , Carballal Guadalupe
Published: March 2007

Guinea pig infection with a highly passaged XJ prototype strain of Junín virus by the intramuscular route (IM) was carried out in order to study viral tropism modification in this host. The neurotropism of this strain was demonstrated by viral isolation, meningitis, and by the presence of Junin antigens as shown by immunofluorescence. These events, not previously observed with the same lower‐passaged strain, revealed the appearance of neurotropism after multiple passages in guinea pigs.
Diagnosis of human parvovirus infection by dot‐blot hybridization using cloned viral DNA
Authors: Anderson M. J. , Jones S. E. , Minson A. C.
Published: March 2007

The human parvovirus can be detected in serum by the immunological techniques of immune electron microscopy (IEM), counterimmunoelectrophoresis (CIE), and radioimmunoassay (RIA). A portion of the genome of this virus has been cloned in pAT153 and used as a 32P‐labelled probe in dot‐blot hybridization assays to detect parvovirus DNA in serum specimens. This test proved a highly sensitive means of detecting virus in microlitre volumes of serum, giving positive results for samples containing 0.5 pg viral DNA, equivalent to 104 virus particles. Unlike CIE and RIA the test is not affected by the presence of parvovirus‐specific antibody in serum specimens, and has permitted virus to be detected in specimens obtained up to 11 days after the onset of clinical symptoms of aplastic crisis.
Detection of human parvovirus using a molecularly cloned probe
Authors: Clewley J. P.
Published: March 2007

Half of the genomic DNA of the human parvovirus (B19) was cloned in the plasmid pBR322. The cloned DNA was used as a molecular probe for the detection of parvovirus in serum by means of a dot hybridization test. In an assay of 26 samples, the dot hybridization test was found to be of comparable sensitivity and to be as rapid as radioimmunoassay for viral antigen detection; it is potentially useful as a diagnostic test.
Use of enzyme‐labeled antigen for the detection of immunoglobulin M and A antibody to herpes simplex virus in serum and cerebrospinal fluid
Authors: van Loon A. M. , van der Logt J. T. M. , Heessen F. W. A. , van der Veen J.
Published: March 2007

A direct enzyme‐linked immunosorbent assay (ELISA that used peroxidase‐labeled antigen) was developed for detection of IgM and IgA antibody to herpes simplex virus (HSV). The assay uses immuno‐affinity‐purified antihuman IgM or IgA antibody‐coated wells of microtiter plates to separate IgM or IgA from other classes of antibody in serum or cerebrospinal fluid (CSF). The presence of specific IgM or IgA is detected by subsequent, consecutive incubation with peroxidase‐labeled antigen and substrate.

HSV antigen was purified by sucrose gradient centrifugation and coupled with peroxidase by the periodate method. By examining sucrose‐gradient‐fractionated sera the assays were shown to be specific for IgM and IgA classes of antibody. None of the sera from patients with Epstein‐Barr virus (n = 20), cytomegalovirus (n = 20), or varicella‐zoster virus (n = 8) infection or with both rheumatoid factor and IgG antibody to HSV (n = 13) reacted positively. Only one out of 78 sera from healthy persons was positive for IgA antibody to HSV, and none for IgM antibody.

All 33 patients with HSV infection developed HSV‐IgA, 22 developed HSV‐IgM. Of the 11 patients with primary infection, all had IgM antibody in their first sera and six had IgA antibody. The corresponding figures for the 22 patients with recurrent infection were five and nine. Furthermore, HSV‐IgA antibody was found in serum and CSF of all five patients with HSV encephalitis in the second week after onset of symptoms, indicating the usefulness of the assay as a nonin‐vasive technique for diagnosing HSV encephalitis.
Antibody response to type‐common and type‐unique epitopes of herpes simplex virus polypeptides
Authors: Bernstein David I. , Bryson Yvonne J. , Lovett Michael A.
Published: March 2007

Herpes simplex virus type 1 (HSV1) and type 2 (HSV2) polypeptides with typecommon and type‐unique epitopes were identified using cross‐adsorbed hyperimmune rabbit sera and Western blotting techniques. Twelve HSVl and fourteen HSV2 polypeptides with type‐specific epitopes were identified. Cross‐adsorbed human sera reacted to a subset of the type‐specific epitopes defined by rabbit sera. Human sera with both HSV1‐ and HSV2‐specific antibodies were identified by their reaction to HSV1 ‐type‐specific epitopes after adsorption with HSV2, and HSV2‐type‐specific epitopes after adsorption with HSV1. Mixing experiments demonstrated the sensitivity of this assay for detecting HSV2‐type‐specific anti‐body in the presence of HSV1 antibody. This technique has proven useful in the study of immune response to individual HSV polypeptides during human HSV infections, while the presence of these type‐unique epitopes on HSV polypeptides allows for the detection of type‐specific antibodies in human sera.
Secretion of polyalbumin receptors in vitro
Authors: Gilja Bal K. , Kasambalides Efthimios J. , Bressler Robert S. , Thung Swan N. , Gerber Michael A.
Published: March 2007

The nature of hepatitis B surface antigen (HBsAg)‐associated receptors for polymerized human serum albumin (pHSA‐R) and their relationship to hepatitis B e antigen (HBeAg) and human serum proteins have not been defined. We studied by radioimmunoassay and by electron microscopy HBsAg‐associated pHSA‐R secreted in vitro by a human hepatocellular carcinoma cell line (PLC/PRF/5) and by mouse 3T3 fibroblasts after transfection with cloned hepatitis B virus (HBV) DNA (4.10 cells). PLC/PRF/5 cells expressed only HBsAg, whereas 4.10 cells secreted also HBeAg. There was no significant difference in the production of HBsAg, HBeAg, and pHSA‐R when the cells were cultured in the presence or absence of fetal calf serum. Secretion of pHSA‐R by the two cell lines for a given amount of HBsAg was equal irrespective of the presence or absence of HBeAg. Supernatants from both cell lines grown in serum‐free medium did not contain any Clq or albumin when tested by immunodiffusion. The ability of a transfected mouse cell line to produce HBsAg with pHSA‐R activity strongly suggests that pHSA‐R is coded by the HBV genome and does not depend on the presence of human serum proteins. In addition, our findings fail to demonstrate any correlation between HBeAg production and pHSA‐R.
Incidence of hepatitis A virus (HAV) infection in rural Liberia
Authors: Prince Alfred M. , Brotman Betsy , Richardson Linda , White Tim , Pollock Nancy , Riddle Janet
Published: March 2007

To provide background for future hepatitis A vaccine trials, sera were collected from 0‐ to 4‐year‐old Liberian infants and their mothers on two occasions an average of 14.75 months apart and tested for antibody to hepatitis A virus (anti‐HAV).

The prevalence of anti‐HAV rose from 2.5% in infants 0–6 months of age to 70% in children 3–4 years of age and did not differ between male and female infants.

The annual incidence of new infections was slightly lower in the first year of life (35%) than in the subsequent 3 years, when it averaged 45%. The presence of HBV infection did not affect the incidence of HAV seroconversion. No clinical hepatitis was recognized in the subjects who seroconverted.

Dual hepatitis A and B virus infection were observed; these were all clinically inapparent.

The extraordinary incidence of HAV infection documented in the present study offers an opportunity for vaccine efficacy trials requiring minimal numbers of subjects.
Antibody response to Epstein‐Barr‐virus‐specific DNase in 13 patients with nasopharyngeal carcinoma in Taiwan: A retrospective study
Authors: Chen Jen‐Yang , Hwang Lu‐Yu , Yang Czau‐Siung , Beasley R. Palmer , Chien Chia‐Siang
Published: March 2007

Serum samples obtained from 13 individuals who were found to have a previous history of or to be suffering from nasopharyngeal carcinoma (NPC) were examined for the presence of antibody to Epstein‐Barr virus (EBV) DNase activity. Significant to high levels of antibody to EBV DNase activity were detected in most serum samples obtained from four patients prior to the diagnosis of NPC. The samples from the other nine patients showed variable levels of antibody. The majority of the samples collected at the remission stage of the disease, especially those from long‐term survivors, contained little or no antibody to the DNase activity. Data presented here suggest that antibody to EBV DNase activity may be a useful marker for the early diagnosis as well as the prognosis of NPC.
Clinical efficacy of a herpes simplex subunit vaccine
Authors: Cappel R. , Sprecher S. , De Cuyper F. , De Braekeleer J.
Published: March 2007

A DNA‐free herpes simplex type 2 subunit vaccine was administered to 18 volunteers without past evidence of herpes simplex type 1 (HSV 1) or herpes simplex type 2 (HSV 2) infection, to 44 patients with severe recurrent genital HSV 2 infection, and to 15 patients with severe oral type 1 HSV recurrences. The vaccine elicited both humoral and cell‐mediated immunity in 97% of the subjects without past HSV infections and boosted significantly the cell‐mediated immunity and antibody titers in almost all the patients with recurrent HSV 1 or HSV 2. The vaccine elicited particularly the production of complement‐dependent cytotoxic antibodies in 96% of the patients with recurrent HSV 2 infections. This might, at least partly, explain the clinical efficacy of the vaccine. Indeed, we observed a significant decrease (t test, p < 0.01) in the attack rate of the recurrences and also a significant shortening of the time needed to complete healing of the lesions (t test, p < 0.01).
Rotavirus prevalence and relationships with climatological factors in gabon, africa
Authors: Sitbon Marc , Lecerf Alain , Garin Yves , Ivanoff Bernard
Published: March 2007

A one‐year study of rotavirus prevalence was carried out using electron microscopy in Gabon, equatorial Africa. One hundred fifty‐six nonhospitalized diarrheic children up to 10 years of age and 115 age‐matched controls were investigated together with diarrheic adults and children over 10 years of age. Rotaviruses were observed only in diarrheic children under 10 years of age (10.2%). Rotaviruses were more prevalent in the dry‐season months (19.2% versus 1.3%, P < .001). Rotaviruses were the most commonly found among all the viral, bacterial, and parasitic agents we detected, with the exception of the coronaviruslike particles, which are the subject of another paper [Sitbon, 1985].
Serum antibodies to the major HSV‐2‐ specified DNA‐binding protein in patients with an acute HSV infection or cervical neoplasia
Authors: Lehtinen Matti , Lehtinen Tuula , Koivisto Vesa , Paavonen Jorma , Leinikki Pauli
Published: March 2007

The major HSV‐2‐specified DNA‐binding protein (ICSP 11/12) was purified from HSV‐infected cells. ELISA and immunoblotting techniques were used to study its antigenicity in HSV‐infected patients and patients with cervical neoplasia and control women. Patients with an acute HSV‐2 infection had clearcut antibody responses to the purified ICSP 11/12 preparation. Determination of the ICSP 11/12 antibodies by ELISA revealed considerably higher serum antibody levels in patients with cervical carcinoma than in the controls.
Epidemic non‐A, non‐B viral hepatitis in algeria: Strong evidence for its spreading by water
Authors: Belabbes El‐Hadj , Bouguermouh Abdelmadjid , Benatallah Ahmed , Illoul Gana
Published: March 2007

A waterborne epidemic of non‐A, non‐B hepatitis occurred in Medea, an Algerian town of 68,000 inhabitants during the autumn and winter of 1980–1981. About 6 weeks before the epidemic, there had been a chlorination failure and an accidental sewage contamination of the river water that supplied large parts of the town. From October, 1980, to January, 1981, 788 cases were notified. This epidemic affected mostly young adults, who usually recovered; however, a high rate of mortality was noted among pregnant women (nine patients, nine deaths). Serological examinations performed on 57 hospitalized patients excluded both hepatitis A and B. The waterborne nature of the epidemic was suggested by a study of the case distribution according to water supplies. The epidemic was severe in the areas supplied by the untreated river water. Few cases were reported in the areas receiving spring water, where chlorination never failed. A mild incidence of infection was noted in the areas supplied by a mixture of untreated river water and chlorinated spring water.
Detection and localization of human papillomavirus DNA in human genital condylomas by in situ hybridization with biotinylated probes
Authors: Beckmann Anna Marie , Myerson David , Daling Janet R. , Kiviat Nancy B. , Fenoglio Cecilia M. , McDougall James K.
Published: March 2007

We have examined the distribution of human papillomavirus (HPV) DNA in paraffin sections of human warts by in situ hybridization with biotin‐labeled DNA probes. Recombinant plasmid DNAs (HPV‐1, −6, −11, −16) were labeled by nick translation with biotinylated deoxyuridine triphosphate. Paraffin sections were hybridized with the probes for 18 h in stringent or nonstringent conditions, and DNA‐DNA hybrids were detected by immunocytochemistry. Paraffin sections of warts were also examined for the presence of HPV capsid antigen with the avidinbiotin peroxidase complex method for immunocytochemistry.

HPV DNA was detected and localized in paraffin sections from a plantar wart, a laryngeal papilloma, and seven anogenital condylomas. The specific HPV type present in each lesion was determined by hybridization under stringent conditions with the homologous DNA probe. The papillomas were found to contain many more cells with replicating virus DNA, as demonstrated by in situ hybridization, than was apparent from the number of cells containing detectable virus antigen. In situ hybridization with biotin‐labeled probes is an effective technique for the identification of HPV infection in routinely collected and processed tissue specimens.
Acute and latent herpes simplex virus neurological disease in mice immunized with purified virus‐specific glycoproteins gB or gD
Authors: Dix Richard D. , Mills John
Published: March 2007

Groups of 5‐week‐old BALB/c mice were immunized intraperitoneally with approximately 10 μg of purified alum‐precipitated glycoprotein gB or gD of either herpes simplex virus type 1 (HSV‐1) or type 2 (HSV‐2) origin. Control mice received injections of alum‐precipitated 1 % bovine serum albumin (BSA). Following a second immunization 4 weeks later, seroconversion was confirmed by demonstrating the presence of glycoprotein‐specific antibody by immune precipitation. All animals were challenged with lethal doses of either HSV‐1 or HSV‐2 by footpad inoculation and assessed for acute virus‐induced neurological disease and the development of ganglionic latency. Whereas 70% of control (BSAimmunized) HSV‐ 1 ‐infected animals developed ascending myelitus and died, 100% of mice immunized with either gB‐1, gB‐2, gD‐1, or gD‐2 antigens remained free of clinical illness and survived HSV‐1 challenge. In contrast, gB‐1 or gB‐2‐immunized mice were not protected against acute HSV‐2‐induced neurological disease and showed a mortality rate of 60–90% (equivalent to that seen in controls), although mean survival times were prolonged. However, significant protection against HSV‐2 challenge was observed with gD‐1 or gD‐2 immunization. When sacral ganglia were removed from surviving mice 9–12 months after virus challenge, latent virus was detected in all gB‐ or gD‐immunized animals, although the extent of latent infection was restricted. These results provide evidence that glycoprotein gD might be superior to glycoprotein gB as an immunogen for the control of acute HSV‐1 and HSV‐2 neurological disease in mice. However, neither glycoprotein prevents ganglionic latency, the source of virus for recurrent herpesvirus infections.
Antibody to Epstein‐Barr virus‐specific DNase as a marker for the early detection of nasopharyngeal carcinoma
Authors: Chen Jen‐Yang , Liu Mei‐Ying , Chen Chien‐Jen , Hsu Mow‐Ming , Tu Shih‐Mien , Lee Hong‐Hsin , Kuo Shiow‐Ling , Yang Czau‐Siung
Published: March 2007

We have previously shown that high levels of antibody to Epstein‐Barr virus (EBV)‐specific DNase may be a useful marker for the early diagnosis of nasopharyngeal carcinoma (NPC). Sera from 3,368 males in an area with a high risk for the development of NPC were examined for the presence of antibody to EBV‐specific DNase activity. Significant levels of the antibody were found in 430 of these sera. As a result, 32 individuals have attended the out‐patient department of otolaryngology for clinical examination. Among them, one individual was found to have a stage II nasopharyngeal carcinoma. This result supports the value of determination of antibody to EBV‐specific DNase as a marker for the early detection of NPC.
Determination of human IgG and IgM class antibodies to West Nile virus by enzyme linked immunosorbent assay (ELISA)
Authors: Feinstein Sabine , Akov Yair , Lachmi Bat‐El , Lehrer Shoshana , Rannon Lotte , Katz David
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) was developed and used for the detection of IgG and IgM antibodies to West Nile virus in human sera. Thirteen paired sera of clinical cases and 24 control sera taken randomly from a blood bank repository were tested. The sera were reacted in microtiter plates coated with PEG‐treated WNV antigen. IgG or IgM antibodies were quantitated by the use of alkaline‐phosphatase‐conjugated anti‐human IgG or IgM antibodies. Of the 24 randomly collected serum samples, 7 were positive in the IgG‐ELISA test. One positive by the IgM‐ELISA was found to contain rheumatoid factor. In 12 of 13 paired sera of clinical cases, IgM as well as IgG antibodies were detected in the second serum sample taken about 3 wk after the onset of clinical signs. The IgM positive sera were screened for rheumatoid factor (RF) on IgG‐coated plates. None of them contained RF. Antibody titers obtained by ELISA showed a good correlation with titers obtained by hemagglutination inhibition, complement fixation, and neutralization tests. The ELISA tests for detection of IgM and IgG antibodies to WNV therefore can replace the other serological methods for epidemiological surveillance and diagnostic purposes.
Acute glandular fever‐like illness in a patient with HTLV‐III antibody
Authors: McCaul T. F. , Tovey G. , Farthing C. F. , Gazzard B. , Zuckerman A. J.
Published: March 2007

A lymph node biopsy obtained from a patient with human T‐cell lymphocytotropic virus III/lymphadenopathy‐associated virus (HTLV‐III/LAV) antibody, presenting with an acute glandular fever‐like illness, was examined by electron microscopy. Numerous pathological changes were present in the biopsy, including hypertrophy of smooth endoplasmic reticulum, intracytoplasmic rod‐like inclusions within the cisternae of endoplasmic reticulum, multivesicular bodies, test‐tube and ringshaped forms, and tubulo‐reticular structures. Intranuclear and intracytoplasmic viral‐like particles measuring 105‐ 120 nm in diameter and small cytoplasmic particles measuring 50–70 nm in diameter were found in some degenerating lymph node cells. These pathological findings may reflect a host cell response to various pathological and viral stimuli resulting from immune deficiency owing to infection with HTLV‐III/LAV.
Evaluation of a rapid passive hemagglutination assay for anti‐rubella antibody: Comparison to hemagglutination inhibition and a vaccine challenge study
Authors: Safford John W. , Abbott Glenda G. , Deimler Colleen M.
Published: March 2007

A rapid passive hemagglutination assay (Rubaquick™) was developed that detects antibody to rubella virus in serum specimens. The test result is read visually after an incubation period of 15–30 minutes. When compared with a hemagglutination inhibition assay, the Rubaquick assay results obtained from 1,470 sera were greater than 99% specific, sensitive, and accurate. Studies of 179 paired serum specimens obtained before and 27 days after rubella vaccination showed that if antibody was detectable by the Rubaquick assay in the prevaccination specimens, the vaccine induced a secondary response consisting of increasing IgG antibody reactivity in the absence of a positive IgM response. In contrast to the positive prevaccination specimens, a negative prevaccination result was associated with IgM antibody in 98 of the 133 postvaccination specimens. Seroconversion was noted in all cases in which the prevaccination specimen was negative by the Rubaquick assay.
Hepatitis B virus in Gizan, Saudi Arabia
Authors: Arya Subhash Chandra , Ashraf Syed Jamaluddin , Parande Chandrakant Mukund , El‐Sayed Mohamad , Sahay Ravi , Ageel Abdul Rahim , Tobeiqi Mohsen Siddiqui
Published: March 2007

The enzyme‐linked immunosorbent assay (ELISA) was used to study the prevalence rates for hepatitis B virus surface antigen (HBsAg), antibody to surface antigen (anti‐HBs), and antibody to core antigen (anti‐HBc) in 724 voluntary donors, students, pregnant women and those seeking treatment for minor ailments in the Gizan area of Saudi Arabia. Tests for hepatitis B e antigen (HBeAg) and e antibody (anti‐HBe) were made in HBsAg positive sera. There was serological evidence of an existing or earlier infection in 337 Saudis (46.5 %), of whom 12.7% were HBsAg carriers, 25.4% were positive for anti‐HBs, and 8.4% were positive only for anti‐HBc. The percentage of HBsAg carriers was 19.9% and 9.3% in males and females, respectively (p < 0.001). The evidence of existing or earlier infection in males (58.7%) was significantly higher than in females (38.7%) (p < 0.001), with no intersex difference in anti‐HBs or anti‐HBc. No difference was observed in the positivity of either of the markers, alone or together, between the cord blood and the female population in the child‐bearing age of 20–39 years. Corresponding to the values in other age groups, there was an overall fall in the number of HBsAg carriers during adolescence as well as in 20–39‐year‐old females. Among the HBsAg carriers, there was no significant difference between the two sexes for HBeAg and anti‐HBe positivity.

The HBsAg carrier rate of 19.9% in males is consistent with the high male dominant prevalence of hepatocellular carcinoma in the Gizan area. Immunization against hepatitis B virus should be considered for all newborns in such hyperendemic areas in the neonatal period.
Changing seroepidemiology of hepatitis A virus infection in Taiwan
Authors: Hsu Hong‐Yuan , Chang Mei‐Hwei , Chen Ding‐Shinn , Lee Ching‐Yun , Sung Juei‐Low
Published: March 2007

Hepatitis A antibody (anti‐HAV) in serum was studied from June to October, 1984, by radioimmunoassay in 647 male and 553 female apparently healthy children under 15 years of age in Taipei City. The prevalence rate of anti‐HAV was 27.0% in infants, decreased to around 1% during the preschool age, then increased and remained around 5% until 11–12 years of age, when another increase was noted, and reached 13.6% among the early teenagers. The agespecific prevalence of anti‐HAV increased with age but differed in three age ranges, which reflected three apparently different calculated annual incidences. Compared with previous studies in Taipei, the results showed a significant reduction in the prevalence of anti‐HAV in almost every age group from 3 to 14 years. This fact probably reflects the marked improvement of hygienic conditions and progress in health education in recent years, which reduced the exposure to HAV infection among young children. The age of primary infection in the children was older than in previous studies, and it is expected that the susceptibility of HAV will extend to early adulthood.
Virus excretion and cell‐mediated immunity during cytomegalovirus infection among healthy infants and children
Authors: Tanaka Ryoko , Tanaka Akira , Oizumi Yoshihumi , Ishida Nakao , Numazaki Yoshio
Published: March 2007

Specific cell‐mediated immunity (CMI) and virus isolation were examined longitudinally to clarify the mechanism of the cessation of virus excretion in inapparent cytomegalovirus (CMV) infection among healthy infants and children. We measured leukocyte procoagulant activity (LPCA) responses to CMV antigen and to a purified protein derivative of tuberculin (PPD), with results expressed as a percentage reduction of recalcification (RC) time. Based on the results in seropositive and seronegative adult control subjects, reductions in RC time of more than 10% were considered indicative of a positive LPCA response. The CMV‐specific LPCA response was negative in all infants shedding the virus, despite the presence of circulating antibodies, but were converted from negative to positive when the virus excretion ceased. This suggests that cessation of the virus excretion in inapparent CMV infection among healthy infants and children probably results from the specific CMI.
Field trial of a Japanese encephalitis diagnostic kit
Authors: Burke Donald S. , Nisalak Ananda , Hoke Charles H.
Published: March 2007

Serum and cerebrospinal fluid (CSF) obtained from patients in rural Thailand during an encephalitis epidemic were assayed with a Japanese encephalitis rapid diagnosis kit. Japanese encephalitis was diagnosed by detection of virus‐specific IgM (JEV IgM) in CSF (1 : 10 dilution) or serum (1 : 100 dilution) with an antibody capture enzyme‐linked immunosorbent assay. Specimens were assayed immediately on site at the provincial hospital and scored by visual examination within 4 h. Each specimen was retested carefully later to accurately determine its activity (units) at a single screening dilution; each was tested also at serial dilutions to determine its end‐point titer. On‐site kit results showed close agreement with subsequent laboratory results for detection and quantitation of JEV IgM and JEV IgG in either serum or CSF. Using the kit on site, admission CSF from 35 (73%) of 48 laboratory‐proven JEV‐infected patients were scored as definitely positive for JEV IgM, while all 17 CSF specimens from non‐JEV infected patients were read as negative (sensitivity 73%, specificity 100%). A rapid and early diagnosis of acute Japanese encephalitis can be accomplished almost anywhere.
Synthesis of measles virus‐specific IgM antibodies and IgM‐class rheumatoid factor in relation to clinical onset of subacute sclerosing panencephalitis
Authors: Ziola B. , Halonen P. , Enders G.
Published: March 2007

Thirty‐seven serum and 37 cerebrospinal fluid (CSF) specimens from 27 patients with subacute sclerosing panencephalitis (SSPE) were tested for measles virus (MV) IgM antibodies and IgM‐class rheumatoid factor (RF) to determine if a temporal relationship exists between SSPE onset and these immunoglobulins. Sensitive solid‐phase radioimmunoassays were used for immunoglobulin detection. Five of six MV IgM‐positive CSF, both MV IgM‐positive sera and six out of eight sera with elevated RF levels were collected within 1 yr of SSPE onset. Also collected during this time were six other sera having high binding in the MV IgM assay, which was not due to MV‐specific IgM antibodies. Two of these sera had as high or higher binding of IgM to a Vero cell control antigen, suggesting involvement of an IgM‐class autoantibody. The other four sera had false‐positive MV IgM assay results due to RF interference. RF interference was dependent on both the titer and avidity of the MV IgG antibodies involved.

Three conclusions can be drawn from these results. First, MV IgM antibodies and elevated RF levles are not markers for acute SSPE, despite the tendency for their synthesis at this stage of the disease. Second, immunoassay analysis of viral IgM antibodies must employ an appropriate control antigen to account for background IgM binding. Finally, even if RF levels are normal or near normal, a false‐positive IgM immunoassay result can still occur if antigen‐specific IgG antibodies in the same sample have the right combination of titer and avidity.
Protection of weanling hamsters from experimental infection with wild‐type parainfluenza virus type 3 (para 3) by cold‐adapted mutants of para 3
Authors: Crookshanks‐Newman Frances K. , Belshe Robert B.
Published: March 2007

Parainfluenza virus type 3 (para 3) was adapted to replicate at 20° C, a nonpermissive temperature for wild‐type (wt) para 3. Serial passage at 20° C resulted in the generation of cold‐adapted (ca) and temperature‐sensitive (ts) mutants. These mutant viruses have been characterized both in vitro and in vivo [Belshe and Hissom (1982): Journal of Medical Virology 10:235–242; Crookshanks and Belshe (1984): Journal of Medical Virology 13:243–249]. We now report the evaluation of three mutants (clone 1150, passaged 12 times in the cold [cp12], clone 1146, passaged 18 times in the cold [cp18], and clone 1328, passaged 45 times in the cold [cp45]) for their ability to protect hamsters from infection by wild‐type para 3. Ether‐anesthetized male syrian hamsters were intranasally vaccinated with either wt para 3 (clone 127) or one of the ca para 3 mutants and on day 28 postvaccination; each animal was intranasally challenged with 105.0 pfu of wt para 3. On days 1, 2, 3, and 4 post‐challenge, 4 to 13 hamsters from each group were sacrificed, and the quantity of para 3 in the nasal turbinates and lungs was determined. Wt virus induced protection from challenge. cp12, cp18, and cp45 reduced the peak titer of wt replication in the lungs by > 100‐fold, tenfold, and tenfold, respectively. The duration of virus replication was shortened also by intranasal vaccination with the mutants. These data give evidence of an inverse relationship between the degree of protection induced by vaccination with coldadapted mutants and the number of passages of the virus in the cold.
Measles IgM antibodies in cerebrospinal fluid and serum in subacute sclerosing panencephalitis
Authors: Chiodi Francesca , Sundqvist Vivi‐Anne , Norrby Erling , Mavra Milica , Link Hans
Published: March 2007

Using a direct enzyme‐linked immunosorbent assay (ELISA), we examined serum and cerebrospinal fluid (CSF) from six patients with subacute sclerosing panencephalitis (SSPE) and control subjects for presence of measles‐virus‐specific IgM antibodies. All samples from the SSPE patients contained demonstrable titers of measles antibodies. The levels of measles IgM antibodies were higher in CSF diluted 1:5 than in serum diluted 1:50, reflecting a local production of IgM antibodies in the central nervous system. Antibody titers remained constant over the course of SSPE in three of the patients followed for three to six months. The IgM ELISA had high sensitivity as well as specificity and was not complicated by false‐positive reactions owing to the presence of rheumatoid factor.
Coxsackie B virus‐specific IgM responses in coronary care unit patients
Authors: Lau Robert C. H.
Published: March 2007

An enzyme‐linked immunosorbent assay (ELISA) test using polyvalent antigens and antisera was used to detect Coxsackie B virus‐specific IgM responses in 329 patients admitted to the Coronary Care Unit, Wellington Hospital, New Zealand over a 12‐month period. The sera of 30 of 153 (19.6%) patients with acute myocardial infarction (AMI), 16 of 98 (18.4%) with chest pain, and 7 of 46 (15.2%) patients with arrhythmia were positive for Coxsackie B virus‐specific IgM. Four of 12 (25%) patients with heart failure were also positive. Over the same period, 178 sex‐ and age‐matched normal blood donors were also studied. Eleven of 178 (6.2%) matched blood donors were positive for Coxsackie B virusspecific IgM. The rates of occurrence of Coxsackie B virus‐specific IgM in patients with AMI and in a group of matched controls showed a significant difference (χ2 = 5.64, p = 0.02).
Hepatitis B virus DNA in leukocytes of patients with hepatitis B virus‐associated liver diseases
Authors: Shen Horng‐Der , Choo Kong‐Bung , Lee Shou‐Dong , Tsai Yang‐Te , Han Shou‐Hwa
Published: March 2007

In order to determine the relationship between hepatitis B virus (HBV) infection of human white blood cells and different forms of HBV‐associated liver diseases, we tested for HBV DNA in the sera and leukocytes of 11 healthy individuals without any serological markers of HBV infection and 91 patients with HBV infection and other gastrointestinal and urinary diseases by dot and Southern blot hybridization. HBV DNA was found in leukocytes of chronic HBV carriers, in acute and chronic hepatitis, and in patients with liver cirrhosis and hepatocellular carcinoma. Between 27 and 50% of individuals in different categories of patients examined were positive for leukocyte HBV DNA. HBV DNA was also detected in the sera of some of these patients but was absent in others. Serum HBV DNA‐positive rates seemed to be highest in hepatitis B e antigen‐positive asymptomatic carriers (8/10, 80%), and tended to drop to lower levels as the disease progressed to liver cirrhosis (0/8) while leukocyte HBV DNA‐positive rates were highest in patients with cirrhosis (4/8, 50%). The results also show that in individuals who were serologically negative for hepatitis B surface antigen (HBsAg) and positive for antibodies to HBsAg and/or HBcAg, HBV DNA was absent in most of the sera (27/28, 96%) but it was present in leukocytes of some of these patients (7/28, 25%). In control experiments with 11 healthy individuals, HBV DNA was not detected in either sera or leukocytes. In all the cases with leukocyte HBV DNA, the HBV DNA molecules were present in free forms with discrete sizes. The exceptions were a case of liver cirrhosis and a case of chronic hepatitis with possible HBV sequence integration into high molecular weight cellular DNA. Since HBV does infect human leukocytes, it may perhaps interfere with the immunological functions of the white blood cells, and thus play an important role in the pathogenesis of HBV‐induced liver disease.
Antigenic homogeneity of adenovirus types 1, 2, 5, and 6
Authors: Kaier Birgit , Wigand Reinhard
Published: March 2007

One hundred twenty strains of adenovirus types 1, 2, 5, and 6 (subgenus C), isolated at various times and places, were tested by neutralization and hemagglutination inhibition with two antisera of each type. Except for one type 5 strain they reacted largely type‐specifically in both tests and neither qualitative nor quantitative differences were observed. Serologically intermediate strains were not found. The antigenic homogeneity is in contrast to the heterogeneity of the genomes of subgenus C adenoviruses.
Inhibitory effect of acyclovir on the growth of inoue‐melnick virus isolated from cerebrospinal fluid
Authors: Nishibe Yoko , Inoue Yukishige K.
Published: March 2007

Inoue‐Melnick virus (IMV) was isolated from the cerebrospinal fluid of patients with chronic neurologic disorders. The effect of acyclovir (ACV), 9‐(2‐hydroxyethoxymethyl)‐guanine, on the growth of IMV was studied in human diploid cell cultures. ACV markedly inhibited the multiplication of all three IMV serotypes in vitro with the same inhibitory effect. By passage of IMV in the presence of ACV in the medium, ACV‐resistant mutants of three IMV serotypes were easily isolated.
Rapid diagnosis of echovirus 33 infection by neutralizing specific IgM antibody
Authors: Pozzetto Bruno , Le Bihan Jean‐Claude , Gaudin Odette‐Germaine
Published: March 2007

In 1982, we isolated 20 strains of echovirus type 33 (EV33) from 18 patients. We studied the humoral response to EV33 of 2,437 subjects from whom at least one serum was available during the same year. In 388 subjects with the neutralizing antibody level at 64 or more, we assayed the EV33 IgM antibodies by seroneutralization test after fractionation of sera by ion exchange chromatography. One hundred ninety‐five subjects (8.0%) had a high titre (≤ 32) of EV33 IgM antibodies, which was considered as evidence of recent infection. The EV33‐positive IgM fractions were assayed against five other enteroviruses. Sixty percent of the IgM fractions did not cross‐react with any of the five serotypes, 8.7% cross‐reacted with at least one serotype but with predominant EV33 IgM response, and 31.3% had an equivalent or greater amount of non‐EV33 IgM antibodies; the type specificity of the assay was directly related to the age of the subjects. These findings suggest that determination of neutralizing specific IgM antibody is a sensitive and rapid test for the diagnosis of enterovirus infections, especially in young people.
Establishment and characterization of a chronic infectious mononucleosislike syndrome in common marmosets
Authors: Emini Emilio A. , Luka Janos , Armstrong Marcy E. , Banker Franklyn S. , Provost Philip J. , Pearson Gary R.
Published: March 2007

Epstein‐Barr virus (EBV) was inoculated into two species of marmosets. Successful infection was established in the majority of the animals of one species, Callithrix jacchus, as evidenced by the development of high, persistent levels of antibody against virus‐specific capsid and early nonstructural proteins. Antibodies also were produced against the major membrane antigen and, in some animals, against EBV nuclear antigen (EBNA) 2 but not against EBNA 1. This is the antibody profile normally noted in individuals with chronic infectious mononucleosis (IM). EBV‐induced lymphoproliferation was not seen, and EBV‐specific proteins were not detected in the peripheral blood lymphocytes of infected animals. Hence, EBV infection in C. jacchus apparently does not generally include extensive B‐cell involvement. However, the marmosets clearly are useful as a model for EBV primary infection and also possibly for chronic IM.
Radioimmunoassay for detection of the snow mountain agent of viral gastroenteritis
Authors: Dolin Raphael , Treanor John J. , Reichman Richard C. , Paul Madore H. , Phillips Michael , Roessner Karen D.
Published: March 2007

The Snow Mountain Agent (SMA) is a Norwalk‐like viral agent of acute gastroenteritis that has been detected only by immune electron microscopy (IEM). We established a solid phase microtiter radioimmunoassay (RIA) for SMA antigen employing pre‐ and post‐challenge sera from volunteer studies as capture antibodies. SMA was detected in 12 of 67 stool samples from volunteers who were ill after oral challenge with SMA. All samples in which virus particles were detected by IEM were positive by RIA, but the RIA was 10–80 times more sensitive than IEM. To detect serum antibody to SMA, a blocking test was developed, employing diarrheal stool containing SMA as a standard source of antigen. Serum antibody rises were detected in eight out of nine volunteers with experimentally induced illness following challenge with SMA, as well as in three out of three naturally occurring cases. A preliminary sero‐epidemiologic survey suggested that infection with SMA was common in the population surveyed. This RIA should permit large scale seroepidemiologic studies of SMA to be carried out, and should also facilitate characterization of this agent.
Homology between HBV‐DNA and a sequence regulating the interferon‐induced antiviral system: Possible mechanism of persistent infection
Authors: Thomas H. C. , Pignatelli M. , Lever A. M. L.
Published: March 2007

Treatment of hepatitis B virus (HBV)‐infected hepatocytes with lymphoblastoid α‐interferon (IFN) leads to an increased expression of the core antigen (HBcAg) and reduced expression of surface antigen (HBsAg). We have identified the presence of a nucleotide sequence at the start of the HBc gene in the hepatitis B virus genome similar to a consensus sequence known to occur upstream from genes induced by IFN in mammalian cells. It is possible that interferon influences the expression of the viral genes because of the presence of this homologous sequence. This sequence in the viral genome may determine the site of integration of the virus and could influence the ability of the cell containing the integrated viral sequence to respond to interferon. This “neutralisation” of the interferon system by viral integration might not only facilitate persistent infection but might also adversely affect response to α‐interferon therapy.