Areversetranscription-PCRassaytodetectviableMycoplasmasynoviaeinpoultryenvironmentalsamplesC.Marois*,C.Savoye,M.Kobisch,I.KempfAgenceFranc¸aisedeSe´curite´SanitairedesAliments(AFSSA),Laboratoired’EtudesetdeRecherchesAvicolesetPorcines,Unite´deMycoplasmologieBacte´riologie,P.O.Box53,F22440Ploufragan,FranceReceived13December2001;receivedinrevisedform31May2002;accepted17June2002AbstractInordertostudyhorizontaltransmissionofMycoplasmasynoviaeanavianpathogen,areversetranscription-polymerasechainreaction(RT-PCR)assaywasdevelopedtodetectviableMycoplasmainenvironment.ThetestwasbasedontheRT-PCRofthe16SribosomalRNA(rRNA)ofMycoplasmagenus.ResultsshowedthatMycoplasma16SrRNAwasstableupto23haftercelldeath.Therefore,thetestalloweddetectionofviableorveryrecently(lessthan23h)deadmycoplasmas.M.synoviaesurvivalinartificiallycontaminatedwater,foodandsoilandintheenvironmentofM.synoviaeexperimentallyinfectedturkeyswasestimatedbycultureandRT-PCR.TheRT-PCRmethodwasthenappliedinanaturallyinfectedlayinghenfarmshowingproblemsofrecurrentmycoplasmosisinthehens.ResultsconfirmedtheusefulnessofRT-PCRincheckingtheefficiencyofbiosecuritymeasuresandinimprovingcleaninganddisinfectionprotocols.#2002ElsevierScienceB.V.Allrightsreserved.Keywords:Mycoplasmasynoviae;Environment;RT-PCR-16SrRNA1.IntroductionAvianmycoplasmosisisafrequentlyreportedinfectiousdiseaseinpoultry.ThetwobacterialspeciesincriminatedareMycoplasmagallisepticumandMycoplasmasynoviae.Theseinfectionscausechronicrespiratorydisease,infectioussinusitisandinfectioussynovitis,resultinginsignificanteconomiclossesbecauseofdecreasedeggproduction,decreasedgrowthandhatchabilityrates,anddowngradingofcarcassesatslaughterbecauseofairsacculitisandarthritislesions(Kleven,1997).VeterinaryMicrobiology89(2002)17–28*Correspondingauthor.Tel.:þ33-2-9601-6281;fax:þ33-2-9601-6273.E-mailaddress:c.marois@ploufragan.afssa.fr(C.Marois).0378-1135/02/$–seefrontmatter#2002ElsevierScienceB.V.Allrightsreserved.PII:S0378-1135(02)00159-1Transmissionofmycoplasma...
