PMID- 29464825
OWN - NLM
STAT- MEDLINE
DCOM- 20180619
LR  - 20191210
IS  - 1365-2672 (Electronic)
IS  - 1364-5072 (Linking)
VI  - 124
IP  - 4
DP  - 2018 Apr
TI  - Performance of pre-RT-qPCR treatments to discriminate infectious human 
      rotaviruses and noroviruses from heat-inactivated viruses: applications of 
      PMA/PMAxx, benzonase and RNase.
PG  - 1008-1016
LID - 10.1111/jam.13737 [doi]
AB  - AIMS: Detection/Quantification of RNA viruses is mostly done by 
      reverse-transcriptase (RT)-(q)PCR, but it does not distinguish between infectious 
      and noninfectious viruses. Our aim was to test, how different pretreatments 
      before RT-qPCR could eliminate positivity originated from external nucleic acids 
      or genomes of damaged particles. METHODS AND RESULTS: Heat-inactivated (80 degrees C for 
      10 min) rotavirus Wa strain and faecal samples containing rotavirus or norovirus 
      were treated with PMA/PMAxx, benzonase or crude extract RNase prior to RT-qPCR. 
      PMA/PMAxx pretreatments were not consistently efficient for RV, although they 
      seemed to work to some extent for heat-inactivated norovirus. Benzonase and RNase 
      provided consistently 2.2-2.8 log(10) reductions in the titre of faecal 
      rotavirus. CONCLUSIONS: All pretreatments need to be further validated for each 
      virus separately, taking into account sample matrix and inactivation conditions. 
      Although none of the pretreatments could completely render inactivated viruses 
      undetectable, RNase worked most consistently for both rota- and norovirus. 
      SIGNIFICANCE AND IMPACT OF THE STUDY: This study sheds light on capacity of the 
      most common pre-RT-qPCR treatments to eliminate damaged, noninfectious 
      rotaviruses and noroviruses after thermal treatment. To our knowledge, this is 
      the first time, when benzonase has been used in this context.
CI  - (c) 2018 The Society for Applied Microbiology.
FAU - Oristo, S
AU  - Oristo S
AUID- ORCID: 0000-0001-6638-8691
AD  - Department of Food Hygiene and Environmental Health, Faculty of Veterinary 
      Medicine, University of Helsinki, Helsinki, Finland.
FAU - Lee, H-J
AU  - Lee HJ
AD  - Department of Food Hygiene and Environmental Health, Faculty of Veterinary 
      Medicine, University of Helsinki, Helsinki, Finland.
FAU - Maunula, L
AU  - Maunula L
AUID- ORCID: 0000-0002-0841-5353
AD  - Department of Food Hygiene and Environmental Health, Faculty of Veterinary 
      Medicine, University of Helsinki, Helsinki, Finland.
LA  - eng
SI  - GENBANK/KT943508
SI  - GENBANK/KT943509
SI  - GENBANK/X81436
SI  - GENBANK/M87661
PT  - Evaluation Study
PT  - Journal Article
DEP - 20180312
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
RN  - EC 3.1.- (Endodeoxyribonucleases)
RN  - EC 3.1.- (Endoribonucleases)
RN  - EC 3.1.- (Ribonucleases)
RN  - EC 3.1.30.2 (Serratia marcescens nuclease)
SB  - IM
MH  - Caliciviridae Infections/*virology
MH  - Discriminant Analysis
MH  - Endodeoxyribonucleases/chemistry
MH  - Endoribonucleases/chemistry
MH  - Hot Temperature
MH  - Humans
MH  - Norovirus/chemistry/genetics/*isolation & purification/physiology
MH  - Reverse Transcriptase Polymerase Chain Reaction/*methods
MH  - Ribonucleases/chemistry
MH  - Rotavirus/chemistry/genetics/*isolation & purification/physiology
MH  - Rotavirus Infections/*virology
MH  - Virus Inactivation
OTO - NOTNLM
OT  - detection
OT  - environmental health
OT  - enzymes
OT  - thermal processes
OT  - viruses
EDAT- 2018/02/22 06:00
MHDA- 2018/06/21 06:00
CRDT- 2018/02/22 06:00
PHST- 2017/06/05 00:00 [received]
PHST- 2018/02/07 00:00 [revised]
PHST- 2018/02/14 00:00 [accepted]
PHST- 2018/02/22 06:00 [pubmed]
PHST- 2018/06/21 06:00 [medline]
PHST- 2018/02/22 06:00 [entrez]
AID - 10.1111/jam.13737 [doi]
PST - ppublish
SO  - J Appl Microbiol. 2018 Apr;124(4):1008-1016. doi: 10.1111/jam.13737. Epub 2018 
      Mar 12.