Rapid detection of Mycoplasma synoviae by loop-mediated isothermal amplification

被引:32
作者
Kursa, Olimpia [1 ]
Wozniakowski, Grzegorz [2 ]
Tomczyk, Grzegorz [1 ]
Sawicka, Anna [1 ]
Minta, Zenon [1 ]
机构
[1] Natl Inst Vet Res, Dept Poultry Dis, PL-24100 Pulawy, Poland
[2] Natl Inst Vet Res, Dept Poultry Viral Dis, PL-24100 Pulawy, Poland
关键词
Mycoplasma synoviae; Loop-mediated isothermal amplification; Rapid detection; POLYMERASE-CHAIN-REACTION; CLINICAL-SAMPLES; GALLISEPTICUM; ASSAY; POULTRY; PCR; FIELD; VLHA; LAMP; ABNORMALITIES;
D O I
10.1007/s00203-014-1063-2
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Mycoplasma synoviae (MS) remains a serious concern in production of poultry and affects world production of chickens and turkeys. Loop-mediated isothermal amplification (LAMP) of DNA has been recently used for the identification of different economically important avian pathogens. The aim of this study was to develop LAMP for simple and inexpensive detection of MS strains in poultry using specifically designed primers targeting hemagglutin A (vlh) gene. The assay was conducted in a water bath for 1 h at 63 A degrees C. The results were visualized after addition of SYBR Green(A (R)) fluorescent dye. LAMP was specific exclusively for MS without cross-reactivity with other Mycoplasma species. The sensitivity of LAMP was determined as 10(-1) CFU/ml and was 1,000 times higher than MS-specific polymerase chain reaction. LAMP assay was conducted on 18 MS field strains to ensure its reliability and usefulness. This is the first report on LAMP development and application for the rapid detection of MS isolated from chickens. This simple method may be applied by diagnostic laboratories without access to expensive equipment.
引用
收藏
页码:319 / 325
页数:7
相关论文
共 36 条
[1]   Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene [J].
Bayatzadeh, Mohammad Ali ;
Pourbakhsh, Seyed Ali ;
Ashtari, Abass ;
Abtin, Ali Reza ;
Abdoshah, Mohammad .
BRITISH POULTRY SCIENCE, 2014, 55 (02) :148-156
[2]   Treatment of Eggshell Abnormalities and Reduced Egg Production Caused by Mycoplasma synoviae Infection [J].
Catania, Salvatore ;
Bilato, Dania ;
Gobbo, Federica ;
Granato, Anna ;
Terregino, Calogero ;
Iob, Luciano ;
Nicholas, Robin A. J. .
AVIAN DISEASES, 2010, 54 (02) :961-964
[3]   Validation of a previously developed quantitative polymerase chain reaction for the detection and quantification of Mycoplasma synoviae in chicken joint specimens [J].
Dijkman, R. ;
Feberwee, A. ;
Landman, W. J. M. .
AVIAN PATHOLOGY, 2013, 42 (02) :100-107
[4]   Development of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Mycoplasma hyorhinis [J].
Du, Gai-Mei ;
Liu, Mao-Jun ;
Wu, Yu-Zi ;
Xiong, Qi-Yan ;
Bai, Fang-Fang ;
Feng, Zhi-Xin ;
Shao, Guo-Qing .
CLINICAL LABORATORY, 2013, 59 (11-12) :1363-1371
[5]   Experimental infection and transmissibility of Mycoplasma synoviae with delayed serologic response in chickens [J].
Ewing, ML ;
Cookson, KC ;
Phillips, RA ;
Turner, KR ;
Kleven, SH .
AVIAN DISEASES, 1998, 42 (02) :230-238
[6]   Induction of eggshell apex abnormalities by Mycoplasma synoviae: field and experimental studies [J].
Feberwee, A. ;
de Wit, J. J. ;
Landman, W. J. M. .
AVIAN PATHOLOGY, 2009, 38 (01) :77-85
[7]   Seroprevalence of Mycoplasma synoviae in Dutch commercial poultry farms [J].
Feberwee, Anneke ;
de Vries, Tjep S. ;
Landman, Wil J. M. .
AVIAN PATHOLOGY, 2008, 37 (06) :629-633
[8]  
Ferguson-Noel N., 2013, Diseases of Poultry, P900
[9]   Apparent eradication of Mycoplasma synoviae in broiler breeders subjected to intensive antibiotic treatment directed to control Escherichia coli [J].
Fiorentin, L ;
Soncini, RA ;
da Costa, JLA ;
Mores, MAZ ;
Trevisol, IM ;
Toda, M ;
Vieira, ND .
AVIAN PATHOLOGY, 2003, 32 (02) :213-216
[10]   A Multiplex real-time PCR for detection of Mycoplasma gallisepticum and Mycoplasma synoviae in clinical samples from Brazilian commercial poultry flocks [J].
Fraga, Aline Padilha ;
de Vargas, Tatiana ;
Ikuta, Nilo ;
Kazantzi Fonseca, Andre Salvador ;
Celmer, Alvaro Jose ;
Marques, Edmundo Kanan ;
Lunge, Vagner Ricardo .
BRAZILIAN JOURNAL OF MICROBIOLOGY, 2013, 44 (02) :505-510