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Novel TaqMan PCR assay for the quantification of Paenibacillus larvae spores in bee-related samples
ID Kušar, Darja (Avtor), ID Papić, Bojan (Avtor), ID Zajc, Urška (Avtor), ID Zdovc, Irena (Avtor), ID Golob, Majda (Avtor), ID Žvokelj, Lucija (Avtor), ID Knific, Tanja (Avtor), ID Avberšek, Jana (Avtor), ID Ocepek, Matjaž (Avtor), ID Pislak, Metka (Avtor)

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Izvleček
Paenibacillus larvae is the causative agent of American foulbrood (AFB), a devastating disease of honeybees. P. larvae spore counts in bee-related samples correlate with the presence of AFB symptoms and may, therefore, be used to identify at-risk colonies. Here, we constructed a TaqMan-based real-time PCR (qPCR) assay targeting a single-copy chromosomal metalloproteinase gene for reliable quantification of P. larvae. The assay was calibrated using digital PCR (dPCR) to allow absolute quantification of P. larvae spores in honey and hive debris samples. The limits of detection and quantification were 8 and 58 spores/g for honey and 188 and 707 spores/mL for hive debris, respectively. To assess the association between AFB clinical symptoms and spore counts, we quantified spores in honey and hive debris samples originating from honeybee colonies with known severity of clinical symptoms. Spore counts in AFB-positive colonies were significantly higher than those in asymptomatic colonies but did not differ significantly with regard to the severity of clinical symptoms. For honey, the average spore germination rate was 0.52% (range = 0.04–6.05%), indicating poor and inconsistent in vitro germination. The newly developed qPCR assay allows reliable detection and quantification of P. larvae in honey and hive debris samples but can also be extended to other sample types.

Jezik:Angleški jezik
Ključne besede:American foulbrood (AFB), Paenibacillus larvae, real-time or quantitative PCR (qPCR), digital PCR (dPCR), plate counting, spore germination rate, honey, hive debris, Polymerase Chain Reaction
Vrsta gradiva:Članek v reviji
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:VF - Veterinarska fakulteta
Status publikacije:Objavljeno
Različica publikacije:Objavljena publikacija
Leto izida:2021
Št. strani:18 str.
Številčenje:Vol. 12, iss. 11, art. 1034
PID:20.500.12556/RUL-133255 Povezava se odpre v novem oknu
UDK:577
ISSN pri članku:2075-4450
DOI:10.3390/insects12111034 Povezava se odpre v novem oknu
COBISS.SI-ID:85522179 Povezava se odpre v novem oknu
Datum objave v RUL:18.11.2021
Število ogledov:1389
Število prenosov:210
Metapodatki:XML DC-XML DC-RDF
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Gradivo je del revije

Naslov:Insects
Skrajšan naslov:Insects
Založnik:MDPI
ISSN:2075-4450
COBISS.SI-ID:519122457 Povezava se odpre v novem oknu

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.
Začetek licenciranja:17.11.2021

Projekti

Financer:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Številka projekta:V4-1804
Naslov:Ugotavljanje poti širjenja hude gnilobe čebelje zalege z genetsko tipizacijo sevov povzročitelja bolezni

Financer:ARRS - Agencija za raziskovalno dejavnost Republike Slovenije
Številka projekta:P4-0092
Naslov:Zdravje živali, okolje in varna hrana

Financer:Drugi - Drug financer ali več financerjev
Program financ.:Republic of Slovenia, Ministry of Agriculture, Forestry, and Food

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