Use of an internal control in a nested-PCR assay for Mycoplasma hyopneumoniae detection and quantification in tracheobronchiolar washings from pigs

2000 ◽  
Vol 14 (6) ◽  
pp. 365-372 ◽  
Author(s):  
E Verdin ◽  
M Kobisch ◽  
JM Bové ◽  
M Garnier ◽  
C Saillard
Keyword(s):  
Internal Control ◽  
Nested Pcr ◽  
Mycoplasma Hyopneumoniae ◽  
Pcr Assay ◽  
Detection And Quantification

Detection of Mycoplasma hyopneumoniae by Air Sampling with a Nested PCR Assay

1998 ◽  
Vol 64 (2) ◽  
pp. 543-548 ◽  
Author(s):  
Katharina D. C. Stärk ◽  
Jacques Nicolet ◽  
Joachim Frey
Keyword(s):  
Nested Pcr ◽  
Gene Segment ◽  
Mycoplasma Hyopneumoniae ◽  
Field Conditions ◽  
Pcr Analysis ◽  
Direct Extraction ◽  
Pcr Assay ◽  
Sampling System ◽  
Respiratory Problems ◽  
One Step

ABSTRACT This article describes the first successful detection of airborneMycoplasma hyopneumoniae under experimental and field conditions with a new nested PCR assay. Air was sampled with polyethersulfone membranes (pore size, 0.2 μm) mounted in filter holders. Filters were processed by dissolution and direct extraction of DNA for PCR analysis. For the PCR, two nested pairs of oligonucleotide primers were designed by using an M. hyopneumoniae-specific DNA sequence of a repeated gene segment. A nested PCR assay was developed and used to analyze samples collected in eight pig houses where respiratory problems had been common. Air was also sampled from a mycoplasma-free herd. The nested PCR was highly specific and 104 times as sensitive as a one-step PCR. Under field conditions, the sampling system was able to detect airborne M. hyopneumoniae on 80% of farms where acute respiratory disease was present. No airborne M. hyopneumoniae was detected on infected farms without acute cases. The chance of successful detection was increased if air was sampled at several locations within a room and at a lower air humidity.

scholarly journals Improved detection of bovine coronavirus N gene in faeces of calves infected naturally by a semi-nested PCR assay and an internal control

2006 ◽  
Vol 131 (2) ◽  
pp. 148-154 ◽  
Author(s):  
Elisabete Takiuchi ◽  
Danilo T. Stipp ◽  
Alice F. Alfieri ◽  
Amauri A. Alfieri
Keyword(s):  
Internal Control ◽  
Nested Pcr ◽  
N Gene ◽  
Pcr Assay ◽  
Bovine Coronavirus

A nested PCR assay for the detection of Mycoplasma hyopneumoniae in tracheobronchiolar washings from pigs

2000 ◽  
Vol 76 (1) ◽  
pp. 31-40 ◽  
Author(s):  
E Verdin ◽  
C Saillard ◽  
A Labbé ◽  
J.M Bové ◽  
M Kobisch
Keyword(s):  
Nested Pcr ◽  
Mycoplasma Hyopneumoniae ◽  
Pcr Assay

A duplex nested-PCR assay for detection of mud crab reovirus and mud crab dicistrovirus-1

2013 ◽  
Vol 20 (4) ◽  
pp. 808-815
Author(s):  
Di ZHANG ◽  
Keng YANG ◽  
Youlu SU ◽  
Juan FENG ◽  
Zhixun GUO
Keyword(s):  
Nested Pcr ◽  
Mud Crab ◽  
Pcr Assay

Development of a hemi-nested PCR assay for the specific detection ofMelissococcus pluton

1998 ◽  
Vol 37 (3) ◽  
pp. 165-174 ◽  
Author(s):  
Steven P Djordjevic ◽  
Kerrie Noone ◽  
Lisa Smith ◽  
Michael A Z Hornitzky
Keyword(s):  
Nested Pcr ◽  
Specific Detection ◽  
Pcr Assay

scholarly journals Neurological disorder in cattle associated with bovine herpesvirus 4

2011 ◽  
Vol 63 (4) ◽  
pp. 828-835 ◽  
Author(s):  
E.A. Costa ◽  
A.C. Vasconcelos ◽  
M.R.Q. Bomfim ◽  
H.B. Amorim ◽  
G.B.L. Lima ◽  
...  
Keyword(s):  
Nested Pcr ◽  
Clinical Course ◽  
Bovine Herpesvirus ◽  
Pcr Assay ◽  
Bovine Herpesvirus 1 ◽  
Herpesvirus Type ◽  
Neurological Signs ◽  
Bovine Herpesvirus 4 ◽  
Herpesvirus 1 ◽  
Herpesvirus 4

A nested PCR assay was used to diagnose bovine encephalitis through herpesviruses including bovine herpesvirus 5 (BHV-5), bovine herpesvirus 1 (BHV-1), Aujeszky's disease virus (SHV-1), and ovine herpesvirus 2 (OHV-2) in 14 fragments of central nervous system (CNS) from cattle that died with neurological signs. In addition, as some samples of bovine herpesvirus type 4 (BHV-4) have been isolated from neural tissue, it was also tested by nested PCR. The cases of encephalitis occurred in isolation at different times of the year and did not present any seasonality. The duration of the clinical course ranged between 1 to 15 days, and in 64.3% of the cases it manifested between 1 to 2 days. The most frequently observed neurological signs were ataxia, recumbency, unsteadiness and inability to stand, opisthotonus, paddling movements, nystagmus and ptyalism. In the nested assay, there was no evidence of: BHV-1, SHV-1 or OHV-2 in the DNA obtained from the CNS in any of the samples. But the presence of BHV-4 was found in all fragments of the CNS in cattle which died presenting neurological signs. Moreover, BHV-5 was found in association with BHV-4 in two of these samples.

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