Nanopore sequencing as a revolutionary diagnostic tool for porcine viral enteric disease complexes identifies porcine kobuvirus as an important enteric virus

Rapid identification of Mycoplasma bovis infections in cattle is a key factor to guide antimicrobial therapy and biosecurity measures. Recently, nanopore sequencing became an affordable diagnostic tool for both clinically relevant viruses and bacteria, but the diagnostic accuracy for M. bovis identification is undocumented. Therefore, in this study nanopore sequencing was compared to rapid identification of M. bovis with MALDI-TOF MS (RIMM), and triplex real-time PCR in a Bayesian latent class model (BLCM) for M. bovis in bronchoalveolar lavage fluid (BALf) obtained from calves. In practice, pooling of samples is often used to save money, but the influence on diagnostic accuracy has not been described for M. bovis . Therefore, a convenience sample of 17 pooled samples containing 5 individual BALf samples per farm was analyzed as well. The results of the pooled samples were compared to the individual samples, to determine sensitivity (Se) and specificity (Sp). The BLCM showed a good Se (77.3%; 95% Credible Interval: 57.8%-92.8%) and high Sp (97.4%; 91.5%-99.7%) for nanopore sequencing compared to RIMM (Se: 93.0%; 76.8%-99.5%, Sp: 91.3; 82.5%-97.0%) and real-time PCR (Se: 94.6%; 89.7%-97.7%, Sp: 86.0%; 76.1-93.6%). Se and Sp of pooled analysis for M. bovis were 85.7% (95% confidence interval: 59.8-111.6%) and 90.0% (71.4-108.6%%) for nanopore sequencing and 100% (100%-100%) and 88.9% (68.4-109.4%) for RIMM, respectively. In conclusion, nanopore sequencing is a rapid, reliable tool for the identification of M. bovis . To reduce costs and increase the chance of M. bovis identification, pooling of 5 samples for nanopore sequencing and RIMM is possible.

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Enteric Disease in Specific-Pathogen-Free Turkey Poults Inoculated with a Small Round Turkey-Origin Enteric Virus

Avian Diseases ◽

10.2307/1591264 ◽

1990 ◽

Vol 34 (3) ◽

pp. 683 ◽

Cited By ~ 7

Author(s):

David E. Swayne ◽

M. Judith Radin ◽

Y. M. Saif

Keyword(s):

Enteric Virus ◽

Specific Pathogen Free ◽

Enteric Disease ◽

Turkey Poults ◽

Specific Pathogen

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Is Porcine Kobuvirus 1 a Typical Diarrhoeic Pathogen of Piglets?

Folia Veterinaria ◽

10.1515/fv-2017-0018 ◽

2017 ◽

Vol 61 (2) ◽

pp. 50-55

Author(s):

H. McFall ◽

Š. Vilček

Keyword(s):

Enteric Virus ◽

Enteric Pathogen ◽

Porcine Kobuvirus ◽

Rt Pcr ◽

Rotavirus A ◽

Pig Farms ◽

Rectal Swabs ◽

Vp6 Gene ◽

3D Gene

AbstractThe objective of this study was to show if porcine kobuvirus 1 (PKV-1) participates in the development of diarrhoea in piglets. The experiments were focused on comparing the occurrence of PKV-1 with the occurrence of rotavirus A (RVA) infection in suckling pigs on Slovak pig farms. A total of 91 rectal swabs of piglets (age < 28 days) were collected from 8 pig farms. RT-PCR was employed to detect PKV-1 through amplification of the 495 bp fragment of the 3D gene using primers KoVF/ KoVR, and RVA was detected through amplification of the 309 bp fragment of the VP6 gene using primers rot3 and rot5. As expected, the detection of RVA in diarrhoeic piglets was 56.8 % (P < 0.01), while only 14.8 % in healthy animals. These results confirm that RVA is one of the main causes of diarrhoea in young piglets. Comparatively, PKV-1 was detected in approximately equal numbers in the same group of both healthy and diarrhoeic pigs, with 74.1 % in healthy animals and 81.1 % in diarrhoeic animals, which was not statistically significant (P < 0.05). The level of co-infection of both viruses was 11.1 % in healthy animals. A portion of 48.6 % (P < 0.01) of diarrhoeic animals were found with RVA and PKV-1 coinfections. The results of this study indicate that while RVA is an enteric virus, PKV-1 cannot confidently be confirmed as an enteric pathogen.

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Scanning Microscopy of Human Intestinal Explants in Organ Culture

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100095248 ◽

1972 ◽

Vol 30 ◽

pp. 396-397

Author(s):

Bruce Wetzel ◽

Robert Buscho ◽

Raphael Dolin

Keyword(s):

Electron Microscopy ◽

Room Temperature ◽

Culture Conditions ◽

Human Fetuses ◽

Enteric Disease ◽

Organ Cultures ◽

Growth Of A Variety ◽

Normal Human ◽

Fetal Intestine ◽

Scanning Electron

It has been reported that explants of human fetal intestine can be maintained in culture for up to 21 days in a viable condition and that these organ cultures support the growth of a variety of known viral agents responsible for enteric disease. Scanning electron microscopy (SEM) has been undertaken on several series of these explants to determine their appearance under routine culture conditions.Fresh specimens of jejunum obtained from normal human fetuses were washed, dissected into l-4mm pieces, and cultured in modified Leibowitz L-15 medium at 34° C as previously described. Serial specimens were fixed each day in 3% glutaraldehyde for 90 minutes at room temperature, rinsed, dehydrated, and dried by the CO2 critical point method in a Denton DCP-1 device. Specimens were attached to aluminum stubs with 3M transfer tape No. 465, and one sample on each stub was carefully rolled along the adhesive such that villi were broken off to expose their interiors.

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The Elicited Language Analysis Procedure

Language Speech and Hearing Services in Schools ◽

10.1044/0161-1461.1301.37 ◽

1982 ◽

Vol 13 (1) ◽

pp. 37-41

Author(s):

Larry J. Mattes

Keyword(s):

Diagnostic Tool ◽

Descriptive Analysis ◽

Quality Measures ◽

Analysis Procedure ◽

Elicited Imitation ◽

Response Quality ◽

Semantic Relationships ◽

Language Analysis

Elicited imitation tasks are frequently used as a diagnostic tool in evaluating children with communication handicaps. This article presents a scoring procedure that can be used to obtain an in-depth descriptive analysis of responses produced on elicited imitation tasks. The Elicited Language Analysis Procedure makes it possible to systematically evaluate responses in terms of both their syntactic and semantic relationships to the stimulus sentences presented by the examiner. Response quality measures are also included in the analysis procedure.

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