<i>Streptococcus equi</i> Detection Polymerase Chain Reaction Assay for Equine Nasopharyngeal and Guttural Pouch Wash Samples

Boyle, A.G.; Rankin, Shelley C.; Duffee, Lauren; Boston, Ray; Wheeler‐Aceto, H.

doi:10.1111/jvim.13808

Cited by 25 publications

(36 citation statements)

References 15 publications

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“…Additionally, there was a single horse with consistent clinical signs, found to be negative on multiple nasal swabs, which was later found to be culture positive from a lymph node aspirate. It is currently recognized that nasal swabs are not as sensitive for recovery of S. equi as guttural pouch lavage or pharyngeal wash samples . It is recommended that in clinical cases with consistent signs the type of sample collected should be adjusted to maximize the chances of recovery.…”

Section: Discussionmentioning

confidence: 99%

Frequency of shedding of respiratory pathogens in horses recently imported to the United States

Smith

Watson

Spier

et al. 2018

Veterinary Internal Medicne

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BackgroundImported horses that have undergone recent long distance transport might represent a serious risk for spreading infectious respiratory pathogens into populations of horses.ObjectiveTo investigate the frequency of shedding of respiratory pathogens in recently imported horses.AnimalsAll imported horses with signed owner consent (n = 167) entering a USDA quarantine for contagious equine metritis from October 2014 to June 2016 were enrolled in the study.MethodsProspective observational study. Enrolled horses had a physical examination performed and nasal secretions collected at the time of entry and subsequently if any horse developed signs of respiratory disease during quarantine. Samples were assayed for equine influenza virus (EIV), equine herpesvirus type‐1, −2, −4, and −5 (EHV‐1, −2, −4, −5), equine rhinitis virus A (ERAV), and B (ERBV) and Streptococcus equi subspecies equi (S. equi) using quantitative PCR (qPCR).ResultsEquine herpesviruses were detected by qPCR in 52% of the study horses including EHV‐2 (28.7%), EHV‐5 (40.7%), EHV‐1 (1.2%), and EHV‐4 (3.0%). Clinical signs were not correlated with being qPCR‐positive for EHV‐4, EHV‐2, or EHV‐5. None of the samples were qPCR‐positive for EIV, ERAV, ERBV, and S. equi. The qPCR assay failed quality control for RNA viruses in 25% (46/167) of samples.Conclusions and Clinical ImportanceClinical signs of respiratory disease were poorly correlated with qPCR positive status for EHV‐2, −4, and −5. The importance of γ‐herpesviruses (EHV‐2 and 5) in respiratory disease is poorly understood. Equine herpesvirus type‐1 or 4 (EHV‐1 or EHV‐4) were detected in 4.2% of horses, which could have serious consequences if shedding animals entered a population of susceptible horses. Biosecurity measures are important when introducing recently imported horses into resident US populations of horses.

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Section: Discussionmentioning

confidence: 99%

Frequency of shedding of respiratory pathogens in horses recently imported to the United States

Smith

Watson

Spier

et al. 2018

Veterinary Internal Medicne

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“…The limit of detection [6] and its use in detection of S. equi in clinical nasopharyngeal wash samples and guttural pouch lavage samples from sick, convalescent, and asymptomatic horses [19] has been published by our laboratory. DNA was extracted from a 1 ml aliquot of the guttural pouch lavage using PrepMan Ultra as described by the manufacturer (Applied Biosystems, Foster City, CA).…”

Section: Methodsmentioning

confidence: 99%

Comparison of nasopharyngeal and guttural pouch specimens to determine the optimal sampling site to detect Streptococcus equi subsp equi carriers by DNA amplification

2017

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Background Streptococcus equi subsp equi (S. equi) is the cause of “equine strangles” which is a highly infectious upper respiratory disease. Detection of S. equi is influenced by site of specimen collection, method of sampling, and type of diagnostic test that is performed. We hypothesized i) that a loop-mediated isothermal amplification (LAMP) assay that targets the S. equi-specific eqbE gene would be more sensitive than a realtime PCR assay that targets the S. equi-specific seeI gene and ii) that LAMP of specimens obtained by guttural pouch lavage (GPL) would be more sensitive than LAMP of nasopharyngeal specimens to identify S. equi carriers.MethodsA nasopharyngeal flocked swab, nasopharyngeal wash, and GPL specimen was collected from 44 convalescent horses and the eqbE LAMP assay was performed. The seeI realtime PCR assay and aerobic culture were also performed on the GPL specimen. Logistic regression was performed to compare sampling sites and test methods (P-values ≤0.05 were considered significant).ResultsOne of 41 nasopharyngeal flocked swabs, 6/38 nasopharyngeal wash and 24/44 GPL specimens were positive by eqbE LAMP. 18/44 GPL specimens were positive by seeI PCR and S. equi was isolated from 4/44 of these specimens. Detection of S. equi DNA was 51 times more likely from the GPL samples than nasopharyngeal samples (OR 51.0, P < 0.0001). When eqbE LAMP GPL samples were positive, it was eight times more likely that the guttural pouch had any abnormality on endoscopy (OR 8.2, P ≤ 0.005), almost 20 times more likely that mild empyema was found (OR 19.7, P ≤ 0.002), and eight times more likely that the SeeI PCR was positive for S. equi DNA (OR 8.1, P ≤ 0.006).ConclusionThis study demonstrates that guttural pouch lavage specimens should be used to detect S. equi and that the eqbE LAMP assay was comparable to the seeI PCR.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-017-0989-4) contains supplementary material, which is available to authorized users.

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“…Polymerase chain reaction for the detection of DNA has been shown to be more sensitive, although more expensive, while supplying same day or next day results (Boyle et al . ,b, , ; Lindahl et al . ; Webb et al .…”

Section: Diagnosticsmentioning

confidence: 99%

“…; Waller ; Boyle et al . , ). Cases of metastatic abscessation may be characterised by anaemia of chronic disease.…”

Section: Diagnosticsmentioning

confidence: 99%

Strangles and its complications

Boyle

2016

Equine Veterinary Education

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Summary Strangles, caused by the Gram‐positive bacteria Streptococcus equi subspecies equi (S. equi), is a highly contagious upper respiratory infection in horses. The infection is transmitted by inhalation or direct contact with mucopurulent discharge from an infected animal, resulting in fever, depression, and submandibular and retropharyngeal lymph node enlargement that can lead to respiratory distress. Complications include secondary cellulitis at external abscessation sites, guttural pouch empyema and its persistence into the carrier state, purpura haemorrhagica, metastatic abscessation, emergency tracheostomies and rarely secondary S. equi pneumonia or myositis. Control of outbreaks requires strict isolation protocols and hygiene measures. Detection methods of the index case and carrier state are constantly being refined to assist in the identification and prevention of disease perpetuation.

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Streptococcus equi Detection Polymerase Chain Reaction Assay for Equine Nasopharyngeal and Guttural Pouch Wash Samples

Cited by 25 publications

References 15 publications

Frequency of shedding of respiratory pathogens in horses recently imported to the United States

Frequency of shedding of respiratory pathogens in horses recently imported to the United States

Comparison of nasopharyngeal and guttural pouch specimens to determine the optimal sampling site to detect Streptococcus equi subsp equi carriers by DNA amplification

Strangles and its complications

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