Avtor/Urednik		Prosenc, Katarina; Kotnik, Antonija; Šubelj, Vesna; Sočan, Maja
Naslov		Primerjava dveh presejalnih metod za odkrivanje influence A, influence B in RSV: ELISA in multiplex RT-PCR
Prevedeni naslov		Comparison of ELISA and multiplex RT-PCR screening assays for detection of influenza A, influenza B and RSV
Tip		članek
Vir		Zdrav Vars
Vol. in št.		Letnik 43, št. 2
Leto izdaje		2004
Obseg		str. 99-103
Jezik		slo
Abstrakt		Acute respiratory tract infections (ARI) are the leading cause of increased morbidity and mortality in winter months. Because of nonspecific clinical symptoms of influenza, laboratory confirmation is essential for surveying influenza spread in the community. Rapid tests, such as enzyme linked immunoassay (ELISA) and multiplex reverse transcription-polymerase chain reaction (RT-PCR), were introduced to replace the time-consuming isolation of influenza and RSV viruses. Multiplex RT-PCR allows for detection of three different antigens during a single test The test was adapted to enable reverse transcription and DNA replication to be processed in the same reaction mix. Throat and nasal swabs were obtained from out-patients and in-patients with ARI. ELISA identified influenza A in 2.5%, influenza 8 in 6.3 % and RSV in 2 % of the 605 clinical samples tested. Detection rates achieved by RT-PCR were 3.6%, 18.6% and 3%, respectively. For isolation in cell culture all ELISA andlor RT-PCR-positive samples were used. Influenza A virus was recovered from two, influenza 8 from 33 and RSV from five samples. RT-PCR was found to be a more sensitive test than ELISA. It improved the rate of virus isolation from RT-PCR-positive samples, which is of utmost importance for typification and sub-typification of influenza viruses. Higher sensitivity of RT-PCR was established in all age groups. RT-PCR yielded better detection results than ELISA, especially in the age group 5 years and over. Despite its higher cost and greater expertise required, RT-PCR offers a significant advantage over ELISA, and is superior to it in influenza screening and RSV detection.
Izvleček		V zimskem času so akutne okužbe dihal vzrok izrazitega porasta obolevnosti in umrljivosti. Med pomembnejšimi povzročitelji so virusi influence in RSV. Zaradi neznačilne klinične slike je potrebno kroženje teh virusov potrditi z virološkimi metodami. Ker je osamitev virusov v celični kulturi zahtevna in dolgotrajna, skušamo prve podatke dobiti s hitrejšimi metodami. Kot presejalni metodi sta bili obravnavni encimsko-imunski test (ELISA) in multipleksna verižna reakcija s polimerazo z reverznim prepisom (RT-PCR). Slednja omogoča odkrivanje več antigenov hkrati. Prilagojena je bila tako, da sta reverzni prepis in pomnoževanje DNK potekala s pripravo ene reakcijske mešanice. Brise žrela in nosu so pri bolnikih s klinično sliko gripi podobne bolezni odvzeli zdravniki, ki sodelujejo v mreži za epidemiološko in virološko spremljanje gripe in zdravniki Klinike za infekcijske bolezni in vročinska stanja v Ljubljani. Obdelanih je bilo 605 vzorcev. Z ELISA je bil dokazan virus influence A v 2,5%, virus influence B v 6,3% in RSV v 2% vzorcev, z RT-PCR pa genom influence A v 3,6%, influence B v 18,6% in RSV v 3% vzorcev. Iz vzorcev, ki so bili pozitivni s katero koli od uporabljenih tehnik, je bilo namnoževanje virusov v celični kulturi za virus influence A uspešno iz 2 vzorcev, za virus influence B iz 33 in za RSV iz 5 vzorcev. Ugotovljeno je bilo, da je presejalna metoda RT-PCR bolj občutljiva kot ELISA in da poveča tudi delež uspešnih osamitev v celičnih kulturah, kar je izrednega pomena za tipizacijo in subtipizacijo virusov influence. Tehnika RT-PCR je bolj občutljiva od tehnike ELISA v vseh starostnih skupinah, razlika pa se v višjih starostnih skupinah povečuje v prid RT-PCR. Posebej je opazno povečanje možnosti odkrivanja virusov influence in RSV z RT-PCR v starostnih skupinah nad 5 let. Kljub večji zahtevnosti in višji ceni se je RT-PCR izkazal za primemejšo presejalno metodo kot ELISA.
Deskriptorji		INFLUENZA INFLUENZA A VIRUS, HUMAN INFLUENZA B VIRUS RESPIRATORY SYNCYTIAL VIRUS, HUMAN ENZYME-LINKED IMMUNOSORBENT ASSAY POLYMERASE CHAIN REACTION