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BOOK REVIEW Substance and seduction Elisa Guerra Doce Department of Prehistoria, Arqueología
Zachary, A. A., Ratner, L. E., Graziani, J. A. és mtsai: Characterization of HLA class I specific antibodies by ELISA using solubilized antigen targets: II. Clinical relevance. Human Immunology, 2001, 62 , 236
The early detection of pregnancy and the determination of fetal numbers have economic benefits in sheep production because of the seasonal breeding patterns where missing a breeding opportunity means the loss of one productive year. The purpose of this study was to evaluate the efficacy of the B6-HRP ELISA for ovine pregnancy-specific protein B (oPSPB) measurement in the detection of pregnancy and estimation of fetal numbers in different sheep breeds. BioPRYN® ELISA assay kit was used for the detection of pregnancy in the experimental animals. Ninety-three ewes of three breeds (British Milksheep – BM, Lacaune – L and Transylvanian Racka – TR), each from three farms in Hungary, were included in the study. BM and L ewes were artificially inseminated (AI). Thirty-five days after AI, all ewes were examined by transabdominal ultrasound. The TR flock was mated naturally over a six-week period. At the end of the mating period, the ewes were similarly examined by ultrasound. Blood samples were taken from all pregnant ewes twice (35 and 65 days after AI), and serum samples were assayed by the BioPRYN test. It can be concluded that the detection of serum PSPB by ELISA is a much easier, safer, less expensive and highly accurate method for the detection of ovine pregnancy. Although some breed-related differences were detectable at 35 and 65 days post breeding, no differences in oPSPB levels were found in pregnant ewes carrying different numbers of fetuses.
Twenty-two serum samples of healthy bitches were tested with the frozen and lyophilised version of the same ELISA kit (Quanticheck, Faculty of Veterinary Science, Budapest, Hungary). Samples were chosen on the basis of their progesterone (P4) concentrations, which were between 1.00 and 20.00 ng/mL. As it is well known, this range has the highest clinical relevance in ovulation diagnosis. Both types of microplates were read at 15-min intervals from the 15th until the 90th minute (min) of incubation, and the results were compared with those of frozen plates at 60 min of incubation as 100 percent. Lyophilised microplates gave on average 18 percent higher results than the frozen version at equal incubation times. The highest difference between lyophilised and frozen samples was observed at 45 and 60 min of incubation. Ninety-four percent of the reaction in the frozen microplate occurred in the first 15 min, and during the subsequent 30 min the reaction seemingly stopped. After the 45th min of incubation, this 94 percent increased to 108 percent in the subsequent 30 min, which remained the final approximate result at the end of the 90 min of incubation. In contrast to the frozen microplate, the measured concentration increased continuously in the lyophilised version and reached the highest level at the 60th min. The results of the lyophilised microplate reached the same level at 30 min of incubation as those of the frozen version at 60 min. In conclusion, a mechanical increase or decrease of the incubation time does not generate a linear change in the test results. This study demonstrated that the results of a series of samples collected from the same bitch cannot be compared if they are measured with different laboratory methods or different ELISA kits.
A commercial ELISA kit (IDEXX MVV/CAEV p28 Ab Screening Test, IDEXX, Montpellier, France) was applied to detect antibodies to SRLVs. This test is an indirect ELISA that uses an immunogenic peptide of a transmembrane protein and a recombinant p28
detection. In small ruminants, serological tests have been used to detect the presence of antibodies against Map, such as the agar gel immunodiffusion (AGID) test, complement fixation tests and the enzyme-linked immunosorbent assay (ELISA) ( Buczinski et al
Changes in ELISA serology are frequently used to determine antibiotic treatment success for Lyme disease in horses. This concept was based upon a previous report showing a marked decline in ELISA values in experimentally infected and antibiotic-treated ponies. Changes in Lyme serology following antibiotic treatment in naturally infected horses have not been reported. The objective of this study was to compare Borrelia ELISA antibody concentrations in naturally exposed horses both before and following antibiotic treatment for Lyme disease. A retrospective study was performed comparing oxytetracycline- or doxycyclinetreated (n = 68) and untreated (n = 183) horses from a single equine practice and their change in Borrelia ELISA values over a similar time period. Antibiotictreated horses had a decline in ELISA values in comparison to control horses (P ≤ 0.05) and untreated horses were twice as likely to have their ELISA values increase (OR = 0.5; 95% C.I. = 0.3–0.9) compared to treated horses. The magnitude of the decline in ELISA units following treatments was small compared to that previously reported in experimentally infected and treated ponies. Field-exposed horses with high Borrelia burgdorferi ELISA values who are treated with either oxytetracycline or doxycycline can be expected to have only a small decline in ELISA values following treatment. Persistently high ELISA titres following appropriate treatments for Lyme disease may not, without appropriate clinical signs, be a reason for more prolonged treatment.
Equine rhinitis B virus 1 (ERBV1), genus Erbovirus, family Picornaviridae , is a pathogen of horses which causes clinical and subclinical infection of the upper respiratory tract in horses. The virus is widespread in European horse populations and the current standard method for the detection of antibody against ERBV1 is by virus neutralisation (VN). VN tests, however, are labour-intensive and time-consuming, require tissue culture facilities, and generally do not provide same-day results. In this study, a protocol for the high-level expression and purification of recombinant virion protein 1 (rVP1) was established using metal-chelate affinity chromatography under denaturing condition. When used as a coating antigen in a prototype enzyme-linked immunosorbent assay (ELISA), denatured rVP1 was recognised by ERBV1 antibody present in horse serum. This finding suggests that denatured rVP1 is a promising candidate for the development of an ELISA to be used in the routine laboratory diagnosis of ERBV1 infection in horses.
A total of 860 serum samples collected at 86 cattle farms in different parts of Hungary were screened for the presence of antibodies to Mycoplasma bovis using an ELISA test with a recombinant M. bovis membrane protein as antigen. Antibodies to M. bovis were detected in sera collected on all farms, and no farms negative for M. bovis were found. In 88.38% of the herds more than 50% of the sampled animals were infected by M. bovis. A total of 82.91% of the animals had antibodies to M. bovis. The proportion of seropositive animals was higher in the older age groups, and a significant difference was seen in the level of seropositivity between young and older age groups. The results show that M. bovis infection is widespread on Hungarian dairy farms, and its prevalence has increased in the recent decade. The high infection rate of Hungarian cattle herds with M. bovis shows that special attention should be paid to evaluating the aetiological role of M. bovis in bovine respiratory disease complex (BRDC) cases because M. bovis has an immunosuppressive effect and can predispose cattle to other respiratory infections, too.
Asprosin levels in the blood sera were determined using a commercial enzyme-linked immunosorbent assay (ELISA) kit (Sunred Bovine Asprosin Kit, Shanghai) and an ELISA reader (Bio Tek Instruments, USA) as described by Ugur and Aydin (2019
