Solid-phase enzyme-linked immunosorbent assay for detection of hepatitis A-specific immunoglobulin M

1979 ◽  
Vol 9 (4) ◽  
pp. 459-465
Author(s):  
S A Locarnini ◽  
A G Coulepis ◽  
A M Stratton ◽  
J Kaldor ◽  
I D Gust
Keyword(s):  
Liver Disease ◽  
Immunosorbent Assay ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dark Urine ◽  
Assay Procedure ◽  
Specific Immunoglobulin

A solid-phase enzyme linked immunosorbent assay was developed for the detection of immunoglobulin M antibody to hepatitis A virus. The system was capable of detecting hepatitis A-specific immunoglobulin M in a single dilution of serum and appears to be a reliable and rapid means of establishing a diagnosis of hepatitis A infection. Specific immunoglobulin M was only detected in patients with serologically confirmed hepatitis A and not in patients with other forms of hepatitis, chronic liver disease, or autoimmune disease. In patients with hepatitis A, specific immunoglobulin M was usually detectable for 6 weeks after the onset of dark urine, and the longest period for which it was present in any patient was 115 days. This enzyme-linked immunosorbent assay is rapid, simple to perform, and does not require complicated equipment. Provided adequate supplies of purified reagents can be obtained, this enzyme-linked immunosorbent assay procedure is likely to simplify hepatitis A serology, because the same antibody-coated plates can be utilized to detect hepatitis A virus, anti-hepatitis A virus, and hepatitis A-specific immunoglobulin M.

scholarly journals Solid-phase enzyme-linked immunosorbent assay for detection of hepatitis A virus

1978 ◽  
Vol 8 (3) ◽  
pp. 277-282 ◽  
Author(s):  
S A Locarnini ◽  
S M Garland ◽  
N I Lehmann ◽  
R C Pringle ◽  
I D Gust
Keyword(s):  
Immunosorbent Assay ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Field Work ◽  
Enzyme Linked Immunosorbent Assay ◽  
Technical Equipment ◽  
Immune Electron Microscopy ◽  
Naked Eye ◽  
Household Contacts

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of hepatitis A virus in human fecal specimens. Investigations with 88 fecal specimens from 77 patients with suspected viral hepatitis and 8 of their household contacts showed that ELISA was as specific and sensitive as radioimmunoassay and almost as sensitive as immune electron microscopy. The ELISA is quick and simple to perform, does not require sophisticated technical equipment, and can be read with the naked eye, making it suitable for field work and rapid diagnosis.

Detection of Hepatitis a Virus in Drinking Water by Enzyme -Immunoassay Using Ultracentrifugation for Virus Concentration

1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger
Keyword(s):  
Membrane Filtration ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Phosphate Buffer Solution ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Concentration ◽  
Buffer Solution ◽  
Solution Ph ◽  
Two Stage

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.

Serodiagnosis of viral hepatitis A by a modified competitive binding radioimmunoassay for immunoglobulin M anti-hepatitis A virus

1979 ◽  
Vol 9 (1) ◽  
pp. 120-127
Author(s):  
D W Bradley ◽  
H A Fields ◽  
K A McCaustland ◽  
J E Maynard ◽  
R H Decker ◽  
...  
Keyword(s):  
Immunoglobulin G ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Protein A ◽  
Competitive Binding ◽  
Immunoglobulin M ◽  
Test Results ◽  
Solid Phase Radioimmunoassay ◽  
Viral Hepatitis A

A competitive binding radioimmunoassay (CBA) for antibody to hepatitis A virus (HAV) was evaluated and compared with a standard solid-phase radioimmunoassay for anti-HAV, CBA was found to be sensitive and specific for the detection of anti-HAV, as demonstrated by the 98% concordance of CBA and solid-phase radioimmunoassay test results. The standard CBA test was modified for the differential detection of acute (immunoglobulin M) and convalescent (immunoglobulin G) anti-HAV by incorporation of a step in which immunoglobulin G anti-HAV was preferentially absorbed with S. aureus cells (protein A). The modified CBA test was shown to be capable of differentiating between acute- and convalescent-phase sera. The modified CBAM test was able to detect immunoglobulin M anti-HAV up to approximately 4 weeks after the onset of illness.

scholarly journals Increasing incidence of hepatitis A in Bangladesh

2012 ◽  
Vol 47 (3) ◽  
pp. 309-312 ◽  
Author(s):  
MZ Amin ◽  
LN Siddique ◽  
MA Slatter ◽  
KK Biswas
Keyword(s):  
Liver Disease ◽  
High Risk ◽  
Chronic Liver Disease ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Age Groups ◽  
Hepatitis E ◽  
Immunoglobulin M ◽  
Chronic Liver ◽  
High Risk Population

Hepatitis A (HAV) infection is caused by the hepatitis A virus which is transmitted through the fecal-oral route. Life long protective antibodies are present after infection. The number of cases of adult hepatitis A has progressively been increasing during the last several decades in Bangladesh. In addition, the pattern of age-specific seroprevalence of anti-HAV has changed with economic growth. The prevalence of anti-HAV in 20-40 year age range has declined rapidly during the last 3 decades. As a result, this age groups has a high risk for HAV infection and clinically overt hepatitis A is increasing in adolescents and adult. The aim of the present study were to assess whether the proportion of adults with acute HAV infection has been increasing over the years and analyze the seroprevalence of immunoglobulin M(IgM) anti- HAV antibodies in young adults below the age of 20 years as well as in cases of chronic liver disease. Sera collected from 530 patients with acute and chronic liver disease attends the Somorita Hospital Ltd. during the previous 2 years and 6 months (Jan. 2008- Jun. 2010) were tested for various serological markers of acute and chronic hepatitis. In addition, 530 normal healthy attendants of the patients above the age of 20 years were tested for IgM anti-HAV as controls. Of 530 patients with acute hepatitis (13.42%) were positive for immunoglobulin M. The patients who were IgM anti-HAV negative were found to be hepatitis B (106 patients), hepatitis C, (10 patients), hepatitis E (150 patients) and unclassified (273 patients). Although the frequency of HAV infection among young adult (< 20 age) had increased (33.33% to 42.35%) in the 2 years and 6 months period, the frequency of HAV infection among adults had also increased (15.38% to 28.13%) during the same period. This study should be helpful for the identification of high risk population for vaccination of hepatitis A. DOI: http://dx.doi.org/10.3329/bjsir.v47i3.13065 Bangladesh J. Sci. Ind. Res. 47(3), 309-312 2012

Evaluation of a Sensitive Reverse Transcription PCR–Enzyme-Linked Immunosorbent Assay for Detection of Hepatitis A Virus in Oysters (Saccostrea glomerata) on the East Coast of the Gulf of Thailand

2014 ◽  
Vol 77 (5) ◽  
pp. 859-863 ◽  
Author(s):  
URAIWAN INTAMASO ◽  
SITTHISAK KETKHUNTHOD
Keyword(s):  
Immunosorbent Assay ◽  
Reverse Transcription ◽  
Hepatitis A ◽  
East Coast ◽  
Hepatitis A Virus ◽  
Detection Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gulf Of Thailand ◽  
Rt Pcr ◽  
The Gulf Of Thailand

Hepatitis A virus (HAV) contamination in food can lead to major health problems. We developed a combination reverse transcription (RT) PCR method plus enzyme-linked immunosorbent assay (ELISA) to detect HAV in fresh oysters harvested along the east coast of the Gulf of Thailand. Viral nucleic acid was extracted via the glycine–arginine–polyethylene glycol method followed by RT-PCR amplification with specifically designed primers against HAV and an ELISA to detect the digoxigenin-labeled RT-PCR products. The ELISA in concert with the RT-PCR protocol further increased the detection sensitivity by 100-fold for the HAV genome and 10-fold in artificially contaminated oysters. The overall sensitivity of the RT-PCR in combination with the ELISA was 31.88 pg and 16 PFU/g, respectively. The ELISA increases the specificity of the RT-PCR assay for detecting naturally occurring HAV in oysters. This combined RT-PCR-ELISA approach is a practical and sensitive method for HAV detection and can be utilized in routine screening for HAV in shellfish.

scholarly journals Immunoglobulin G Immunity to Hepatitis A Virus in Liver Transplant Candidates: A Serosurvey from Iran

2021 ◽  
Vol 21 (2) ◽  
Author(s):  
Kamran B Lankarani ◽  
Behnam Honarvar ◽  
Mohammad Hassan Zahedroozegar ◽  
Alireza Dehghan ◽  
Mohammad Reza Rouhezamin ◽  
...  
Keyword(s):  
Liver Disease ◽  
Liver Transplant ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Multivariable Analysis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Waterpipe Smoking ◽  
Abdominal Sonography ◽  
End Stage Liver Disease ◽  
End Stage

Background: Becoming infected with hepatitis A virus (HAV) is deadlier in patients with end-stage liver disease. Objectives: This study aimed to determine the seroprevalence of chronic immunity to HAV in liver transplant (LT) candidates to determine whether HAV vaccination is necessary for them or not. Methods: This cross-sectional study was conducted on adult LT candidates who were referred to the LT center of Shiraz, Iran. The patients were interviewed for filling the data collection forms. These forms consisted of demographic information, medical backgrounds, etiology of chronic liver disease, a model for end-stage liver disease (MELD) score, laboratory findings, and abdominal sonography report. Furthermore, a 3-cc blood sample was obtained from each patient, and anti-HAV IgG was detected by Enzyme-linked Immunosorbent assay (ELISA) using standard Diapro kits. Univariable and multivariable data analyses were performed using SPSS version 20. A P-value of less than 0.05 was considered the significant cutoff in regression analysis. Results: A total of 291 patients with a mean age of 47.73 ± 12.9 years were recruited in this study of whom, 197 (67.7%) patients were males, 237 (81.4%) were married, 229 (78.7%) were educated lower than 12 years, 250 (85.9%) were living in urban areas, and (221) 75.9% had access to sanitary water in their living area. anti-HAV IgG was detected in 269 (92.4%, 95% CI: 89.4 - 95.4%) patients. Multivariable analysis showed that lower knowledge of hepatitis A transmission routes (OR: 11.9, 95% CI: 1.39 - 101.8, P = 0.024), no waterpipe smoking (OR: 9.5, 95% CI: 1.6 - 55.5, P = 0.014), and older age (OR: 1.12, 95% CI: 1 - 1.24, P = 0.03) were the main predictors of HAV immunity, in sequence. Conclusions: Most LT candidates are HAV IgG positive, but due to the growing number of LT candidates and high mortality of HAV in non-immune cases, LT candidates should be checked for HAV IgG, especially younger or waterpipe smoking patients who are less immune. Also, all non-immune patients should be vaccinated against HAV, if possible.

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