Evaluation of the ELISA and comparison to the complement fixation test and radial immunodiffusion enzyme assay for detection of antibodies against Mycoplasma hyopneumoniae in swine serum

Bereiter, M.; Young, Theresa F.; Joo, Heesoo; Ross, Richard F.

doi:10.1016/0378-1135(90)90075-7

Cited by 74 publications

(63 citation statements)

References 8 publications

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“…Strain 232 of M. hyopneumoniae was isolated from a pig infected with strain 11 (4,30) and has been commonly used to study virulence and vaccination regimens in the United States (49,54,55). Chromosomal DNA was isolated by the method of Artiushin et al (1), and two libraries were constructed.…”

Section: Methodsmentioning

confidence: 99%

The Genome Sequence of Mycoplasma hyopneumoniae Strain 232, the Agent of Swine Mycoplasmosis

et al. 2004

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Section: Methodsmentioning

confidence: 99%

The Genome Sequence of Mycoplasma hyopneumoniae Strain 232, the Agent of Swine Mycoplasmosis

et al. 2004

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“…From the existing serologic tests, complement fixation (CF), indirect hemagglutination, and enzymelinked immunosorbent assay (ELISA), the ELISA is the most sensitive, although CF has been shown to detect antibodies earlier (2 weeks after infection). 1,4 The appearance of antibodies to M. hyopneumoniae, however, can be delayed by as much as 8 weeks after infection, making serologic interpretation less useful for the detection of recent infections in young pigs. 16 In late infections, it is common to find clinically affected pigs that reach slaughter while still seronegative.…”

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confidence: 99%

Application of a Nested Polymerase Chain Reaction Assay to Detect Mycoplasma Hyopneumoniae from Nasal Swabs

1999

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Abstract. The porcine respiratory disease complex (PRDC) is an increasingly important cause of decreased swine productivity and is characterized by slow growth, decreased feed efficiency, anorexia, cough, and dyspnea. Mycoplasma hyopneumoniae is among the most prevalent and important infectious agents associated with PRDC. Understanding of mycoplasmal pneumonia has been hindered by inadequate diagnostic methods. Many of the currently available tests are relatively insensitive or nonspecific when used in a diagnostic laboratory setting or are too costly or difficult for routine diagnostic use. Several polymerase chain reaction (PCR) assays have been described, but they are not sensitive enough to detect the microorganisms in live pigs, from either nasal or tracheal swabs. A nested PCR using 2 species-specific sets of primers from the 16S ribosomal DNA gave positive results with as little as 80 microorganisms and did not cross-react with other mycoplasma species or with other microorganisms commonly found in the respiratory tract of pigs. This assay was better suited for detection of M. hyopneumoniae from nasal swabs than was conventional PCR. Nasal swab samples were taken at different time periods following experimental challenge of 10 susceptible pigs. Only 2 of the 55 swabs examined gave a positive result with conventional PCR, whereas 30 of the 55 swabs gave a positive result using the nested PCR. Twenty of 40 (50%) nasal swabs from pigs experiencing a respiratory disease outbreak where M. hyopneumoniae had been diagnosed also gave a positive result with the nested PCR. To confirm that the amplified product was specific, 4 nested PCR products were purified, sequences were determined and aligned, and they were confirmed to be from M. hyopneumoniae.

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“…3 Several attempts have been made to overcome this specificity problem by using different methods of antigen preparation. Several workers have used purified Tween 20 extracts of M. hyopneumoniae and found improved specificity.…”

Section: Discussionmentioning

confidence: 99%

Differential Serologic Response to Mycoplasma Ovipneumoniae and Mycoplasma Arginini in Lambs Affected with Chronic Respiratory Disease

et al. 1999

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Abstract. An enzyme-linked immunoabsorbent assay (ELISA) was used to evaluate the levels of antibodies to Mycoplasma ovipneumoniae and M. arginini in lambs with chronic respiratory disease. Sera were obtained from lambs in several flocks at various stages of the clinical disease and tested with sodium dodecyl sulfate (SDS)-treated M. ovipneumoniae and M. arginini whole cells and a crude capsular extract of M. ovipneumoniae as the antigens. There were low levels of antibody to M. ovipneumoniae in flocks sampled at the early stages of infection, whereas increased levels of antibody were present in lambs from flocks that had apparently recovered from the clinical disease. Slowly rising titers of circulating antibodies to M. ovipneumoniae were confirmed by sequential bleeding of lambs during the course of the clinical disease. However, antibody levels of M. arginini were more likely to increase earlier in the disease process. There was significant cross-reactivity between the 2 SDS-treated antigens in both the ELISA test and western immunoblotting. In contrast, the crude capsular extract was specific for detecting antibodies to M. ovipneumoniae.Mycoplasma ovipneumoniae (MO) is one of the most commonly isolated microorganisms from sheep with respiratory disease worldwide. 4 Recently in this laboratory, this microorganism and M. arginini (MA) have been routinely recovered from young lambs with a respiratory disease that has been termed the ''coughing syndrome.'' The condition is associated with a severe paroxysmal cough, leading to rectal prolapses. This syndrome is widespread in the midwestern states of the USA, with morbidity and severity variable among flocks. The disease is chronic and persists for several weeks in most affected lambs. There is some evidence that the extended persistence of the MO organism in the respiratory tract of these lambs and the chronic nature of the disease may be due to failure of the immune system to generate protective immunity. 10 Reasons for that failure are not known, but marked variation in the MO organisms isolated from sheep 7 and expression of a polysaccharide capsule by the organisms are recognized. 11 Because of its simplicity the enzyme-linked immunosorbent assay (ELISA) has been extensively used in detecting antibodies to Mycoplasma species. However, concerns exist about its specificity because of crossreactions among several Mycoplasma species. 3 Several attempts have been made to overcome such specificity problems by using different methods of antigen preparation. 6,12 Solubilized whole cell antigens of MO have been used, and cross-reactivity problems were noted with these preparations. 5 Consequently, a crude capsular material was extracted from MO for use as a diagnostic antigen, and the specificity of this reagent was confirmed in ELISA and western immunoblotting procedures. Humoral response to MO could be distinguished from humoral response to MA when measured with crude capsular material extracted from the MO organisms. Levels of antibodies to MO and MA antigens in th...

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Evaluation of the ELISA and comparison to the complement fixation test and radial immunodiffusion enzyme assay for detection of antibodies against Mycoplasma hyopneumoniae in swine serum

Cited by 74 publications

References 8 publications

The Genome Sequence of Mycoplasma hyopneumoniae Strain 232, the Agent of Swine Mycoplasmosis

The Genome Sequence of Mycoplasma hyopneumoniae Strain 232, the Agent of Swine Mycoplasmosis

Application of a Nested Polymerase Chain Reaction Assay to Detect Mycoplasma Hyopneumoniae from Nasal Swabs

Differential Serologic Response to Mycoplasma Ovipneumoniae and Mycoplasma Arginini in Lambs Affected with Chronic Respiratory Disease

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