Enzyme-Linked Immunosorbent Assay (Elisa) for Detection of Human Reovirus-Like Agent of Infantile Gastroenteritis

Yolken, Robert H.; Kim, Hyun Wha; Clem, Thomas R.; Wyatt, Richard G.; Kalica, Anthony R.; Chanock, Robert M.; Kapikian, Albert Z.

doi:10.1016/s0140-6736(77)90951-5

Cited by 310 publications

(117 citation statements)

References 15 publications

Supporting

Mentioning

112

Contrasting

Unclassified

Order By: Relevance

“…Infection with Type 1 was asymptomatic in 45 per cent of cases, whereas every Type 2 infection was associated with gastroenteritis. This postulated greater virulence of type of human rotavirus is similar to that previously observed for Type 1 poliovirus, which exceeds the Type 2 and Type 3 polioviruses in neuropathogenicity 7 One of the fascinating and currently unexplained aspects of rotavirus epidemiology is the frequent occurrence of asymptomatic infection during the newborn period." , 19 Our analysis of eight such strains from England indicates that each was Type 2 -the same type that was associated with gastroenteritis that led to admission of older children to the same bospital.…”

Section: Discussionsupporting

confidence: 72%

Epidemiology of Human Rotavirus Types 1 and 2 as Studied by Enzyme-Linked Immunosorbent Assay

et al. 1978

View full text Add to dashboard Cite

To determine the relative importance of two known serotypes of human rotavirus, we developed an enzyme-linked immunosorbent assay to differentiate serotype-specific rotavirus antigen and antibody. Using this technic, we studied the epidemiology of the two serotypes in acute gastroenteritis. Seventy-seven per cent of 414 rotavirus isolates were Type 2, and the remainder were Type 1. The serotype distribution was similar in specimens from children in Washington D.C., and other parts of the world. Sero-R OTAVIRUS is an important cause of gastroenteritis of infants and children in many parts of the world.'' 2 Because the virus does not grow effi-ci,ently in tissue culture, conventional neutralization "methods cannot be used to examine its serotypic variations.''' Recently, two human rotavirus serotypes were distinguished by complement fixation,' immune electron microscopy' and immunofluorescence. 6 The latter two methods are not practical for serotyping of large numbers of rotavirus-positive specimens whereas complement fixation is not sensitive enough for use with low-titered preparations. We recently described the technic of enzyme-linked immunosorbent assay for the detection of rotavirus antigen and antibody.'"" This assay is similar in design to radioimmunoassay but uses an enzyme instead of a radioactive isotope to quantitate binding of immunoglobulin. We modified the basic assay for detection of rotavirus serotypes and used this new procedure to study the epidemiology of these viruses. MATERIALS AND METHODS Specimens'Stool specimens containing rotavirus were collected from 278 in-il. nts and children with gastroenteritis living in the Washington, D.C., area. From January, 1974, to October, 1976, only hospitalized patients were studied, whereas from November, 1976, to May, 1978, specimens from both inpatients and outpatients were available. This population has been described in detail.' In addition, 136 rotaviruscontaining specimens were available from children with and without gastroenteritis from other localities throughout the world. Included were rotavirus-positive stool specimens obtained

show abstract

Section: Discussionsupporting

confidence: 72%

Epidemiology of Human Rotavirus Types 1 and 2 as Studied by Enzyme-Linked Immunosorbent Assay

et al. 1978

View full text Add to dashboard Cite

show abstract

“…Because infants and children are particularly susceptible to complications such as dehydration that are secondary to the acute enteritis caused by these agents, methods of rapid diagnosis are clinically important. ELISA and RIA have been recently used for this purpose (10,11,14). As reported here, USERIA is an improved method of rapid diagnosis that is several orders of magnitude more sensitive than ELISA and RIA.…”

Section: Discussionmentioning

confidence: 97%

Ultrasensitive enzymatic radioimmunoassay: application to detection of cholera toxin and rotavirus.

Harris¹,

Yolken²,

Krokan³

et al. 1979

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Rotavirus and enterotoxin-producing bacteria are major causes of diarrheal disease in humans. A method of rapid diagnosis, ultrasensitive enzymatic radioimmunoassay, has been developed to quantitatively detect cholera toxin and rotavirus. The method uses features of both enzyme-linked immunosorbent assay and radioimmunoassay; however, the sensitivity of the assay is 100-to 1000-fold more sensitive than the two parent assays. Ultrasensitive enzymatic radioimmunoassay should also be useful in measuring other biologically important agents such as drugs and hormones.In recent years, the capability to detect small quantities of biologically important molecules has been greatly expanded by the development of radioimmunoassays (RIA) (1, 2). These assays have been especially useful for the detection of circulating substances, such as hormones, drugs, and infectious agents. However, RIA have certain practical limitations. The short half-life of the isotopes used limits the shelf life of the reagents. In addition, the need for y-emitting isotopes subjects the users of RIA to a radiation hazard. Enzyme immunoassays (also known as enzyme-linked immunosorbent assays or ELISA) have been developed in an attempt to overcome those problems (3-5). ELISA is similar in design to solid-phase RIA except that an enzyme is used as the immunoglobulin marker instead of a 'y-emitting isotope. This enzyme-antibody conjugate is bound to the solid phase by a series of antibody-antigen reactions and essentially converts the substrate to products with a visible yellow color, which can be measured spectrophotometrically. The fact that a single molecule of enzyme is capable of reacting with a large number of substrate molecules provides for amplification and, thus, a high degree of sensitivity (3, 6). Because stable enzymes such as alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] can be used, ELISA also has the advantage of using reagents with a long shelf life.In practice, however, the sensitivity of ELISA systems has not significantly exceeded that of RIA (7). In order to combine the advantages of both RIA and ELISA, we developed a practical ultrasensitive enzymatic radioimmunoassay (USERIA) for the detection of antigens such as human rotavinus (an important cause of infantile gastroenteritis) and cholera toxin. These assay systems are a 1000-fold more sensitive than both RIA and ELISA for the detection of these agents. MATERIALS AND METHODSReagents.[3H]AMP (generally 3H-labeled, 15 Ci/mmol, New England Nuclear; 1 Ci = 3.7 X 10O°becquerels) was purified by column chromatography with DEAE-Sephadex (A-25; Pharmacia) in a stepwise manner as follows: (i)

show abstract

“…The method cannot be relied upon to detect virus concentrations under a million per gram of faeces (Madeley, 1979). Specimens were subsequently screened for rotavirus by ELISA (Yolken et al 1977) using the 'Rotazyme' kit supplied by Abbott Laboratories. Salmonella, shigella and campylobacter species, and in infants enteropathogenic Escherichia coli, were sought by standard methods.…”

Section: Methodsmentioning

confidence: 99%

Virus diarrhoea associated with pale fatty faeces

Thomas

Luton

Mortimer

1981

J. Hyg.

View full text Add to dashboard Cite

SUMMARYSteatorrhoea was a significant feature in an outbreak of rotavirus gastroenteritis which affected adults and infants in hospital. Fat globules or fatty acid crystals were obvious by light microscopy (LM) in faeces from 14 of 25 patients examined. Ten of the fatty stools and two of the remainder were very pale. By electron microscopy (EM) a rotavirus was seen in 11 of the 14 fatty faeces and in only two of 11 specimens without visible fat.In a further study of pale or fatty faeces 20 such specimens sent for laboratory examination from patients not involved in the hospital outbreak were compared microbiologically with a similar number which were neither pale nor fatty. Viruses were found by EM in 11 (55 %) of the pale or fatty stools; eight rotaviruses, two astroviruses and an uncultivable adenovirus were seen; one further patient had acute jaundice. In contrast, no viruses were seen by EM in the twenty specimens which were normally pigmented and without evident fat.Steatorrhoea was significantly associated with rotavirus infection of the alimentary tract which usually presented as a fatty enteritis. We conclude that rotaviruses certainly, and other viruses possibly, can impede both the digestion of fat and the pigmentation of the faeces. Inspection and LM of faeces are easy. In acute enteritis a fatty or pale stool is an indication for virological examination.

show abstract

Enzyme-Linked Immunosorbent Assay (Elisa) for Detection of Human Reovirus-Like Agent of Infantile Gastroenteritis

Cited by 310 publications

References 15 publications

Epidemiology of Human Rotavirus Types 1 and 2 as Studied by Enzyme-Linked Immunosorbent Assay

Epidemiology of Human Rotavirus Types 1 and 2 as Studied by Enzyme-Linked Immunosorbent Assay

Ultrasensitive enzymatic radioimmunoassay: application to detection of cholera toxin and rotavirus.

Virus diarrhoea associated with pale fatty faeces

Contact Info

Product

Resources

About