Use of a multiplex-polymerase chain reaction for detection of Salmonella and Chlamydophila psittaci from caged birds
Abstract
In this study, development of a multiplex-PCR (m-PCR) assay for identification of Salmonellosis and Psittacosis in caged birds was aimed. DNAs extracted from Salmonella spp. strains from the collection of our department and Salmonella spp. isolates from caged birds in addition to the extracted DNA samples belong to two different Chlamydophila psittaci clinical strains (strain 6BC and a strain isolated from a parakeet), constituted the material of the study. A m-PCR assay was developed which use iroBF and iroBR primers to specifically amplify 606 bp sequence of iroB gene of Salmonella spp. DNA in the same reaction mix and with the same conditions, CpsiA and CpsiB primers were used to amplify 300 bp sequence of pmp gene of C. psittaci. The specificity and the sensitivity of the assay were determined by using positive controls (Salmonella and Chlamydia DNA samples). We believe that the newly developed m-PCR assay is a fast, reliable and an economic assay, which has the potential for direct identification of Salmonella spp. and C. psittaci from the clinical samples
References
- Ali A, Reynolds DL (2000): A multiplex reverse transcription-polymerase chain reaction assay for Newcastle disease virus and avian pneumovirus (Colorado strain). Avian Dis, 44, 938-943.
- Al-Soud WA, Radstrom P (2000): Effects of amplification facilitators on diagnostic PCR in the presence of blood, feces, and meat. J Clin Microbiol, 38, 4463–4470.
- Baumler AJ, Heffron F, Reisbrodt R (1997): Rapid detection of Salmonella enterica with primers specific for iroB. J Clin Microbiol, 35, 1224-1230.
- Carli KT, Unal CB, Caner V, Eyigor A (2001): Detection of salmonella in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis. J Clin Microbiol, 39, 1871- 1876.
- Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE (2000): Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev, 13, 559-570.
- Everett KDE, Hornung LJ, Andersen AA (1999): Rapid detection of the Chlamydiaceae and other families in the order Chlamydiales: three PCR tests. J Clin Microbiol, 37, 575-580.
- Eyigor A, Carli KT (2003): Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes. Avian Dis, 47, 380-386.
- Grimes JE (1987): Zoonoses acquired from pet birds. Vet Clin North Am Small Anim Pract, 17, 209-218.
- Henegariu O, Heerema NA, Dlouhy SR, Vance GH, Vogt PH (1997): Multiplex PCR: critical parameters and step-by-step protocol. Biotechniques, 23, 504–511.
- Herring AJ (1993): Typing Chlamydia psittaci-a review of methods and recent findings. Br Vet J, 149, 455-475.
- Hewinson RG, Griffiths PC, Bevan BJ, Kirwan SES, Field ME, Woodward MJ, Dawson M (1997): Detection of Chlamydia psittaci DNA in avian clinical samples by polymerase chain reaction. Vet Microbiol, 54, 155-166.
- Hewinson RG, Rankin SES, Bevan BJ, Field M, Woodward MJ (1991): Detection of Chlamydia psittaci from avian field samples using the PCR. Vet Rec, 128, 129-130.
- Hong Y, Liu T, Hofacre C, Maier M, White DG, Ayers S, Wang L, Maurer JJ (2003): A restriction fragment length polymorphism-based polymerase chain reaction as an alternative to serotyping for identifying Salmonella serotypes. Avian Dis, 47, 387-395.
- Laroucau K, Souriau A, Rodolakis A (2001): Improved sensitivity of PCR for Chlamydophila using pmp genes. Vet Microbiol 82, 155-164.
- Malorny B, Hoorfar J (2005): Toward standardization of diagnostic PCR testing of fecal samples: lessons from the detection of Salmonellae in pigs. J Clin Microbiol, 43, 3033-3037.
- McElnea CL, Cross GM (1999): Methods of detection of Chlamydia psittaci in domesticated and wild birds. Aust Vet J, 77, 516-521.
- Messmer TO, Skelton SK, Moroney JF, Daugharty H, Fields BS (1997): Application of a nested, multiplex PCR to psittacosis outbreaks. J Clin Microbiol, 35, 2043-2046.
- Moroney JF, Guevara R, Iverson C, Chen FM, Skelton SK, Messmer TO, Plikaytis B, Williams PO, Blake P, Butler JC (1998): Detection of chlamydiosis in a shipment of pet birds, leading to recognition of an outbreak of clinically mild psittacosis in humans. Clin Infect Dis, 26, 1425-1429.
- Olsen B, Persson K, Broholm KA (1998): PCR detection of Chlamydia psittaci in faecal samples from passerine birds in Sweden. Epidemiol Infect, 121, 481-484.
- Orosz SE, Chengappa MM, Oyster RA, Morris PJ, Trock S, Altekruse S (1992): Salmonella enteritidis infection in two species of Psittaciformes. Avian Dis, 36, 766-769.
- Pang Y, Wang H, Girshick T, Xie Z, Khan MI (2002): Development and application of a multiplex polymerase chain reaction for avian respiratory agents. Avian Dis, 46, 691-699.
- Riyaz-Ul-Hassan S, Verma V, Qazi GN (2004): Rapid detection of Salmonella by polymerase chain reaction. Mol Cell Probes, 18, 333-339.
- Ryan TP (1985): Salmonellosis in caged birds. Mod Vet Pract, 66, 383-385.
- Singh DV (2003): Hexaplex PCR for rapid detection of virulence factors. Expert Rev Mol Diagn, 3, 781-784.
- Wilson IG (1997): Inhibition and facilitation of nucleic acid amplification. Appl Environ Microbiol, 63, 3741- 3751.
Kafes kuşlarında Salmonella ve Chlamydophila psittaci’nin saptanmasında multiplex-polimeraz zincirreaksiyonu tekniğinin kullanılması
Abstract
Bu çalışmada, Salmonellosis ve Psittacosis’in tanısında kullanılmak üzere bir multipleks-polimeraz zincir reaksiyonu (m-PZR) tekniği geliştirilmesi amaçlandı. Ankara Üniversitesi Veteriner Fakültesi Mikrobiyoloji Anabilim Dalı suş koleksiyonunda yer alan Salmonella suşları, kafes kuşu orijinli Salmonella izolatları ve bunlardan izole edilen DNA örnekleri dışında yurt dışından sağlanan ve iki farklı Chlamydophila psittaci klinik suşundan izole edilmiş DNA örnekleri çalışma materyalini oluşturdu. Çalışmada, Salmonella türlerinde iroB genini kodlayan sekansın 606 bp’lik porsiyonunu spesifik olarak amplifiye eden iroBF ve iroBR primer çifti ile Chlamydophila psittaci’nin pmp genlerinin 300 bp’lik sekansını amplifiye eden CpsiA ve CpsiB primer çiftinin aynı reaksiyon karışımı ve şartlarında çalışacağı bir m-PZR tekniği geliştirildi. Pozitif kontrol Chlamydophila psittaci DNA’larının dışında, kendi laboratuvar izolatları ve suşlarından izole edilen DNA örnekleri kullanılarak tekniğin spesifite ve sensitivitesi de belirlendi. Sonuç olarak, klinik materyallerden direkt tanı potansiyeli de bulunan tekniğin kafes kuşlarında Salmonella spp. ve C. psittaci’nin tanısında hızlı, güvenilir ve ekonomik bir teknik olduğu düşünülmektedir
References
- Ali A, Reynolds DL (2000): A multiplex reverse transcription-polymerase chain reaction assay for Newcastle disease virus and avian pneumovirus (Colorado strain). Avian Dis, 44, 938-943.
- Al-Soud WA, Radstrom P (2000): Effects of amplification facilitators on diagnostic PCR in the presence of blood, feces, and meat. J Clin Microbiol, 38, 4463–4470.
- Baumler AJ, Heffron F, Reisbrodt R (1997): Rapid detection of Salmonella enterica with primers specific for iroB. J Clin Microbiol, 35, 1224-1230.
- Carli KT, Unal CB, Caner V, Eyigor A (2001): Detection of salmonella in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis. J Clin Microbiol, 39, 1871- 1876.
- Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE (2000): Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev, 13, 559-570.
- Everett KDE, Hornung LJ, Andersen AA (1999): Rapid detection of the Chlamydiaceae and other families in the order Chlamydiales: three PCR tests. J Clin Microbiol, 37, 575-580.
- Eyigor A, Carli KT (2003): Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes. Avian Dis, 47, 380-386.
- Grimes JE (1987): Zoonoses acquired from pet birds. Vet Clin North Am Small Anim Pract, 17, 209-218.
- Henegariu O, Heerema NA, Dlouhy SR, Vance GH, Vogt PH (1997): Multiplex PCR: critical parameters and step-by-step protocol. Biotechniques, 23, 504–511.
- Herring AJ (1993): Typing Chlamydia psittaci-a review of methods and recent findings. Br Vet J, 149, 455-475.
- Hewinson RG, Griffiths PC, Bevan BJ, Kirwan SES, Field ME, Woodward MJ, Dawson M (1997): Detection of Chlamydia psittaci DNA in avian clinical samples by polymerase chain reaction. Vet Microbiol, 54, 155-166.
- Hewinson RG, Rankin SES, Bevan BJ, Field M, Woodward MJ (1991): Detection of Chlamydia psittaci from avian field samples using the PCR. Vet Rec, 128, 129-130.
- Hong Y, Liu T, Hofacre C, Maier M, White DG, Ayers S, Wang L, Maurer JJ (2003): A restriction fragment length polymorphism-based polymerase chain reaction as an alternative to serotyping for identifying Salmonella serotypes. Avian Dis, 47, 387-395.
- Laroucau K, Souriau A, Rodolakis A (2001): Improved sensitivity of PCR for Chlamydophila using pmp genes. Vet Microbiol 82, 155-164.
- Malorny B, Hoorfar J (2005): Toward standardization of diagnostic PCR testing of fecal samples: lessons from the detection of Salmonellae in pigs. J Clin Microbiol, 43, 3033-3037.
- McElnea CL, Cross GM (1999): Methods of detection of Chlamydia psittaci in domesticated and wild birds. Aust Vet J, 77, 516-521.
- Messmer TO, Skelton SK, Moroney JF, Daugharty H, Fields BS (1997): Application of a nested, multiplex PCR to psittacosis outbreaks. J Clin Microbiol, 35, 2043-2046.
- Moroney JF, Guevara R, Iverson C, Chen FM, Skelton SK, Messmer TO, Plikaytis B, Williams PO, Blake P, Butler JC (1998): Detection of chlamydiosis in a shipment of pet birds, leading to recognition of an outbreak of clinically mild psittacosis in humans. Clin Infect Dis, 26, 1425-1429.
- Olsen B, Persson K, Broholm KA (1998): PCR detection of Chlamydia psittaci in faecal samples from passerine birds in Sweden. Epidemiol Infect, 121, 481-484.
- Orosz SE, Chengappa MM, Oyster RA, Morris PJ, Trock S, Altekruse S (1992): Salmonella enteritidis infection in two species of Psittaciformes. Avian Dis, 36, 766-769.
- Pang Y, Wang H, Girshick T, Xie Z, Khan MI (2002): Development and application of a multiplex polymerase chain reaction for avian respiratory agents. Avian Dis, 46, 691-699.
- Riyaz-Ul-Hassan S, Verma V, Qazi GN (2004): Rapid detection of Salmonella by polymerase chain reaction. Mol Cell Probes, 18, 333-339.
- Ryan TP (1985): Salmonellosis in caged birds. Mod Vet Pract, 66, 383-385.
- Singh DV (2003): Hexaplex PCR for rapid detection of virulence factors. Expert Rev Mol Diagn, 3, 781-784.
- Wilson IG (1997): Inhibition and facilitation of nucleic acid amplification. Appl Environ Microbiol, 63, 3741- 3751.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA75CB73CE |
| Journal Section | Research Article |
| Authors | |
| Publication Date | December 1, 2009 |
| Published in Issue | Year 2009 |