The Effect of Pooling Sera on the Detection of Avian Pneumovirus Antibodies using an Enzyme-Linked Immunosorbent Assay Test

Abstract

Abstract. Pooling of samples is a cost-effective approach to estimate disease prevalence and to identify infected individuals. The objective of this study was to evaluate the use of serum pools for the detection of avian pneumovirus infection in turkey flocks by enzyme-linked immunosorbent assay, so that a minimum number of tests can be performed without compromising the sensitivity and specificity of the test. A total of 900 field samples were tested; 20 samples from each of 45 flocks. All samples were tested individually followed by pool testing in groups of 3, 4, 5, and 7 samples each. The number of positive pools for a given pool size was positively associated with the number of positive samples. In a separate experiment, the effect of dilution was examined by pooling 1 positive sample with different numbers of negative samples to form pools of sizes 2-7. These laboratory results were analyzed and integrated into a simulation model aimed at evaluating costefficient testing procedures. The probability of detecting an infected flock depended on prevalence of infection, size of serum pool, and the cutoff value used for optical density difference. At a theoretical prevalence of 20%, the probability of detecting an infected flock was 0.93 and 0.86 for a pool of 2 and 7, respectively. The probability of detecting positive flocks increased with increased prevalence and decreased cutoff. Pooling of samples represented a significant reduction in the cost of testing, suggesting that pooling is more advantageous and cost effective than testing individual samples.

Pilot Investigation on the Presence of Anti-Hepatitis E Virus (HEV) Antibodies in Piglet Processing Fluids

“…The use of pooled samples allows testing of a large number of animals, reducing time and cost. However, a major limitation to using pools as diagnostic tools is the dilution of positive samples below the limit of detection, resulting in misdiagnosed false negatives [32,33]. Therefore, in the current study, the dilution effect when multiple samples are pooled was investigated.…”

Assessing the performance of multiplexed tandem PCR for the diagnosis of pathogenic genotypes of Theileria orientalis using pooled blood samples from cattle

“…1). Previously, [21] had also showed that the dilution of serum (pools of three, four, five or seven) for the detection of avian pneumovirus antibodies using an enzyme-linked immunosorbent assay led to falsenegative results. Similarly, Chatterjee et al [38] investigated the detection of hepatitis B virus in human blood donors using nucleic acid testing (NAT), and found that the pooling of blood samples led to an increase in false-negative results, particularly for samples with a low virus load.…”

“…Indeed, diagnostic specificity and sensitivity depend on a number of factors, including inhibitory factors in the sample matrix, pool size and robustness of the assay [21][22], and each of these aspects need to be critically assessed. Therefore, in the present study, we evaluated the performance of the MT-PCR assay for the diagnosis of the two pathogenic genotypes (chitose and ikeda) of T. orientalis in cattle using individual and pooled blood samples.…”

Rapid detection of Grapevine leafroll-associated virus type 3 using a reverse transcription loop-mediated amplification method

“…Pooling samples allows for a larger number of samples to be tested quickly and more cheaply (Muñoz-Zanzi et al, 2000;Wells et al,2003;Maherchandani et al, 2004;Brinkhof et al, 2007) .However one of the greatest disadvantages of pooling samples is that it can result in a loss sensitivity of the diagnostic tests (Brinkhof et al, 2007). In the case of ELISA, the pooling of samples can also result in unacceptably high OD of samples which limits the amount of samples which can be tested together (Brinkhof et al, 2007).…”