The development of indirect and sandwich ELISA-based detection methods for the detection of Campylobacter jejuni using monoclonal and polyclonal antibody preparations

Abstract This study aimed to evaluate the effects of feeding avian-derived polyclonal antibody preparation (PAP; CAMAS, Inc.) against Streptococcus bovis, Fusobacterium necrophorum, and lipopolysaccharides (40, 35, and 25% of the preparation, respectively) on growth performance of beef cattle during the backgrounding phase. From d 0 to 56, Angus crossbreed heifers (n = 80; 360 ± 60 kg of BW; 470 ± 26 d of age) and steers (n = 20; 386 ± 65 kg of BW; 465 ± 30 d of age) were blocked by BW and randomly assigned to 1 of 16 concrete-floored pens (108 m2), equipped with 2 GrowSafe (GrowSafe Systems Ltd., Airdrie, Alberta, Canada) feed bunks each. Animals received a common ad libitum diet (76% TDN, 15.9% CP, DM basis) with the addition of 1 (PAP1), 3 (PAP3), or 0 g (CON) of PAP per d. Feed intake was recorded daily and BW were obtained on d -1, 0, 14, 28, 42, 55, and 56, to assess changes in BW, ADG, DMI, and G:F. Based upon orthogonal contrasts (CON vs. PAP1, and PAP1 vs. PAP3), BW and ADG on d 14, and DMI from 0 to 28, and 0 to 42 were greater for PAP1 vs. CON (P ≤ 0.03), whereas PAP3 animals were intermediate (P ≥ 0.20). No differences in final BW, DMI, and ADG from d 0 to 56 were detected among treatments (P ≥ 0.22). In conclusion, feeding 1g of polyclonal antibody preparations against Streptococcus bovis, Fusobacterium necrophorum, and lipopolysaccharides in a backgrounding diet, improved growth performance in the first 14 d of feeding suggesting that feeding these PAP for longer than 14 d may not be necessary. The effects on subsequent feedlot performance when using PAP should be evaluated in future studies

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Comparison of Molecular Subtyping and Antimicrobial Resistance Detection Methods Used in a Large Multistate Outbreak of Extensively Drug-Resistant Campylobacter jejuni Infections Linked to Pet Store Puppies

Journal of Clinical Microbiology ◽

10.1128/jcm.00771-20 ◽

2020 ◽

Vol 58 (10) ◽

Author(s):

Lavin A. Joseph ◽

Louise K. Francois Watkins ◽

Jessica Chen ◽

Kaitlin A. Tagg ◽

Christy Bennett ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Campylobacter Jejuni ◽

Multilocus Sequence Typing ◽

The United States ◽

Sequence Type ◽

Outbreak Detection ◽

Detection Methods ◽

Drug Resistant ◽

Molecular Subtyping ◽

Content Type

ABSTRACT Campylobacter jejuni is a leading cause of enteric bacterial illness in the United States. Traditional molecular subtyping methods, such as pulsed-field gel electrophoresis (PFGE) and 7-gene multilocus sequence typing (MLST), provided limited resolution to adequately identify C. jejuni outbreaks and separate out sporadic isolates during outbreak investigations. Whole-genome sequencing (WGS) has emerged as a powerful tool for C. jejuni outbreak detection. In this investigation, 45 human and 11 puppy isolates obtained during a 2016–2018 outbreak linked to pet store puppies were sequenced. Core genome multilocus sequence typing (cgMLST) and high-quality single nucleotide polymorphism (hqSNP) analysis of the sequence data separated the isolates into the same two clades containing minor within-clade differences; however, cgMLST analysis does not require selection of an appropriate reference genome, making the method preferable to hqSNP analysis for Campylobacter surveillance and cluster detection. The isolates were classified as sequence type 2109 (ST2109)—a rarely seen MLST sequence type. PFGE was performed on 38 human and 10 puppy isolates; PFGE patterns did not reliably predict clustering by cgMLST analysis. Genetic detection of antimicrobial resistance determinants predicted that all outbreak-associated isolates would be resistant to six drug classes. Traditional antimicrobial susceptibility testing (AST) confirmed a high correlation between genotypic and phenotypic antimicrobial resistance determinations. WGS analysis linked C. jejuni isolates in humans and pet store puppies even when canine exposure information was unknown, aiding the epidemiological investigation during the outbreak. WGS data were also used to quickly identify the highly drug-resistant profile of these outbreak-associated C. jejuni isolates.

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Greenhouse Comparison of Two Detection Methods for Potato virus YN-Wi at Four Potato Growth Stages

Plant Health Progress ◽

10.1094/php-11-17-0072-rs ◽

2018 ◽

Vol 19 (1) ◽

pp. 71-75 ◽

Cited By ~ 1

Author(s):

Abby Beissinger ◽

Debra Ann Inglis

Keyword(s):

Potato Virus ◽

Detection Method ◽

Sandwich Elisa ◽

Potato Production ◽

Detection Methods ◽

Growth Stages ◽

Veinal Necrosis ◽

Leaf Drop ◽

Potato Growth ◽

Common Laboratory

Potato virus Y (PVY) causes significant crop and monetary losses. Owing to the prevalence of newly emerging strains of PVY such as PVYN-Wi, which often cause asymptomatic to mild reactions on certain potato cultivars, accurate tools are required to detect the virus in potato production. This study compared the sensitivity of a rapid field detection method (immunostrips) with a common laboratory detection method (triple antibody sandwich ELISA) on cultivar Chieftain, grown under isolated conditions in a greenhouse and mechanically inoculated with PVYN-Wi, at four potato growth stages (emergence, preflower, postflower, and senescence). Plants inoculated at emergence displayed severe symptoms of mosaic, veinal necrosis, and leaf drop. Plants inoculated at preflower, postflower, and senescence had veinal necrosis but low or no incidence of mosaic and leaf drop. Overall, few or no tuber symptoms were observed, but a trend of lower tuber yield occurred for emergence-inoculated plants. Low variability in PVYN-Wi detection occurred in both tests for emergence-inoculated plants, whereas those inoculated at preflower and postflower had more variability. Because symptom expression may differ depending on the growth stage when a plant becomes infected, these variations should be heeded with either detection method when collecting samples for PVY testing.

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AABH (aspartyl [asparaginyl] β-hydroxylase) as a serum biomarker for breast cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.5_suppl.2 ◽

2020 ◽

Vol 38 (5_suppl) ◽

pp. 2-2

Author(s):

Kiarash Moshiri

Keyword(s):

Breast Cancer ◽

Sandwich Elisa ◽

Survival Rates ◽

Elevated Serum ◽

Disease Recurrence ◽

The United States ◽

Develop Breast Cancer ◽

Detection Methods ◽

Great Promise ◽

Primary Tumors

2 Background: Breast cancer accounts for 27% of female cancers. One in every eight women will develop breast cancer in their lifetime. More than 220,000 new cases of breast cancer will be diagnosed in the United States in 2020. Five-year survival rates for breast cancer are 98% for localized disease, 83% for regionally spread and 26% for distantly spread cancer. However, 25% of all metastases occur more than 5 years after the initial diagnosis and thus survival rates decrease as time goes out to 10 years due to disease recurrence. Current detection methods rely on imaging modalities, including mammography and MRI. No serum bio-markers have been approved for either breast cancer screening or monitoring for disease recurrence, yet. In asymptomatic women in remission from breast cancer the guidelines of the major oncologic organizations only suggest annual mammography for detection of new primary tumors. Methods: We have investigated the utility of aspartyl (asparaginyl) β-hydroxylase (AABH) as a serum bio-marker for cancer. AABH has been detected by immunohistochemical staining (IHC) in a broad range of cancers including breast cancer. It has been detected by IHC in > 99% of tumor specimens tested (n > 2000) but is absent in adjacent non-affected tissue, and in tissue samples from non-affected individuals. This led to the development of a sandwich ELISA that reliably measures AABH in serum. Results: In the current study we have utilized the assay to quantify AABH levels in the sera of patients diagnosed with Breast Cancer compared to women free of disease. Increased levels of AABH were found in the serum of 99% of patients with breast cancer (n = 189). In women not known to have cancer, AABH was essentially undetectable in serum (n = 65, specificity = 96%). Serum from several women currently in remission subsequent to treatment for breast cancer were also analyzed and found to be negative for AABH. Conclusions: Thus, measurement of serum AABH levels has great promise as a diagnostic tool for breast cancer with potential application in monitoring for disease recurrence. Elevated serum AABH in conjunction with mammography and MRI may greatly facilitate earlier diagnosis of both primary and recurrent Breast Cancer.

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Evaluation of a polyclonal antibody based sandwich ELISA for the detection of faecal antigens in Schistosoma spindale infection in bovines

Journal of Parasitic Diseases ◽

10.1007/s12639-012-0129-9 ◽

2012 ◽

Author(s):

G. S. Sreenivasa Murthy ◽

Placid E. D’Souza ◽

K. Shrikrishna Isloor

Keyword(s):

Polyclonal Antibody ◽

Sandwich Elisa

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132 Measurement of ficus benjamina antigen using polyclonal antibody sandwich ELISA

Journal of Allergy and Clinical Immunology ◽

10.1016/s0091-6749(96)80350-2 ◽

1996 ◽

Vol 97 (1) ◽

pp. 215-215 ◽

Cited By ~ 4

Author(s):

M FALLER ◽

F DEBLAY ◽

E LETT ◽

A VEROT ◽

J BESSOT ◽

...

Keyword(s):

Polyclonal Antibody ◽

Sandwich Elisa ◽

Ficus Benjamina

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Lactate Dehydrogenase Polyclonal Antibody Sandwich ELISA for Determination of Endpoint Heating Temperatures of Ground Beef

Journal of Food Protection ◽

10.4315/0362-028x-59.1.51 ◽

1996 ◽

Vol 59 (1) ◽

pp. 51-55 ◽

Cited By ~ 7

Author(s):

CHENG-HSIN WANG ◽

MOHAMED M. ABOUZIED ◽

JAMES J. PESTKA ◽

DENISE M. SMITH

Keyword(s):

Lactate Dehydrogenase ◽

Polyclonal Antibody ◽

Heating Temperature ◽

Polyclonal Antibodies ◽

Sandwich Elisa ◽

Ground Beef ◽

Enzyme Linked Immunosorbent Assay ◽

Protein Marker ◽

Beef Patties

An enzyme-linked immunosorbent assay (ELISA) for the protein marker lactate dehydrogenase (LDH) was developed to determine endpoint heating temperature (EPT) of ground beef. Ground beef was placed in 10 by 75 mm tubes and heated to temperatures between 62 and 74°C at 2°C intervals. Electrophoresis and immunoblotting of meat extracts indicated a decrease in the intensity of the LDH band as temperature was increased. Polyclonal antibodies (PAb) against bovine muscle LDH were produced and a sandwich ELISA devised using biotinylated PAb as the detecting antibody. The LDH content decreased (P < 0.05) in ground beef heated between 66 and 74°C and was less than 4 μg/g of meat at the recommended minimum endpoint of 69°C. The ELISA was used to estimate the EPT of commercially cooked beef patties.

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Aspartyl (asparaginyl) β-hydroxylase (AABH), a serum biomarker for lung cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.5_suppl.45 ◽

2020 ◽

Vol 38 (5_suppl) ◽

pp. 45-45

Author(s):

Kiarash Moshiri

Keyword(s):

Lung Cancer ◽

Sandwich Elisa ◽

Elevated Serum ◽

Ct Scanning ◽

The United States ◽

Serum Biomarker ◽

Detection Methods ◽

Great Promise ◽

Lower Sensitivity ◽

Serological Screening

45 Background: There are no serum biomarkers currently approved for the detection of lung cancer, a leading cause of cancer associated mortality world wide. More than 180,000 cases of lung cancer will be diagnosed in the United States in 2020 by other means such as by X-ray and CT scanning methods, which have inherently lower sensitivity and higher cost when compared generally to serological methods. While the five year survival for lung cancer is 15%, a survival rate of 50% can be achieved when detection is made early in individuals with localized cancer. Current detection methods, however, enable such detection in only about 18% of cases overall. Methods: A prospective serum biomarker Aspartyl (Asparaginyl) ß Hydroxylase (AABH), has been previously found to be elevated by immunohistochemical staining (IHC) in a broad range of cancers, including lung cancer. AABH was detected in > 99% of tumor specimens tested (n > 1000) but absent in adjacent tissue. The present study introduces a double monoclonal sandwich ELISA which provides detection and comparative quantification of AABH in serum of lung cancer patients vs. normal, and high-risk controls such as cigarette smokers without cancer. This is relevant since 87% of lung cancers are attributable to cigarette smoking, and associative parallels can be seen with recent reductions in rates of smoking. Results: Increased levels of serum AABH were found in 99% of patients with lung cancer (n = 192). Serum AABH was found to be undetectable in individuals not known to have cancer (n = 129, specificity = 93%). In patients with lung cancer, AABH was detectable at all stages. In a population of 50 smokers not known to have cancer, the mean serum AABH level was 0 ng/ml with 90% specificity. Conclusions: The AABH serum ELISA therefore has great promise as an additional diagnostic tool for lung cancer having the practicality and cost effectiveness of conventional serological screening. Elevated serum AABH in conjunction with CT scanning may greatly facilitate earlier diagnosis of lung cancer at a stage in which cure rates are significantly higher and thus may contribute to increased patient survival.

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