PMID- 20651038
OWN - NLM
STAT- MEDLINE
DCOM- 20101104
LR  - 20230529
IS  - 1473-5644 (Electronic)
IS  - 0022-2615 (Print)
IS  - 0022-2615 (Linking)
VI  - 59
IP  - Pt 11
DP  - 2010 Nov
TI  - Investigation of human haemotropic Mycoplasma infections using a novel generic 
      haemoplasma qPCR assay on blood samples and blood smears.
PG  - 1285-1292
LID - 10.1099/jmm.0.021691-0 [doi]
AB  - The aim of this study was to develop quantitative real-time (q)PCR assays to 
      detect all known haemoplasma species, and a human housekeeping gene in order to 
      demonstrate both successful DNA extraction from clinical samples and to test for 
      sample inhibition, and to apply these qPCRs to human blood samples and blood 
      smears. Sensitive and specific generic haemoplasma qPCR assays were developed to 
      amplify haemoplasma species, as well as human glyceraldehyde-3-phosphate 
      dehydrogenase (GAPDH) as an internal amplification control. An optimized 
      technique for extracting DNA from stained blood smears was also developed. These 
      methods were applied to anonymized blood samples obtained from 100 human 
      immunodeficiency virus (HIV)-infected South Africans and 920 UK patients 
      undergoing haematological examination, and to 15 blood smears recruited from 
      previous studies describing human haemoplasmosis. Human GAPDH levels were 
      acceptable in all but three of the blood samples and all but two of the blood 
      smears. The latter could have arisen due to DNA degradation due to the old age 
      (over 35 years) of these smears. Haemoplasma infection was found in one 
      HIV-infected South African, but the species could not be characterized due to the 
      very low levels of DNA present. This report describes novel extraction and qPCR 
      methodologies for haemoplasma screening. Previously reported human haemoplasmosis 
      based on cytological diagnosis alone should be viewed with caution.
FAU - Tasker, Severine
AU  - Tasker S
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Peters, Iain R
AU  - Peters IR
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Mumford, Andrew D
AU  - Mumford AD
AD  - Bristol Heart Institute, Bristol Royal Infirmary, Bristol BS2 8HW, UK.
FAU - Day, Michael J
AU  - Day MJ
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Gruffydd-Jones, Timothy J
AU  - Gruffydd-Jones TJ
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Day, Sarinder
AU  - Day S
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Pretorius, Anne-Marie
AU  - Pretorius AM
AD  - University of the Free State, Bloemfontein, South Africa.
FAU - Birtles, Richard J
AU  - Birtles RJ
AD  - University of Liverpool, Chester High Road, Chester CH64 7TE, UK.
FAU - Helps, Chris R
AU  - Helps CR
AD  - School of Veterinary Sciences, University of Bristol, Langford, Bristol BS40 5DU, 
      UK.
FAU - Neimark, Harold
AU  - Neimark H
AD  - State University of New York, Brooklyn, NY, USA.
LA  - eng
GR  - WT_/Wellcome Trust/United Kingdom
GR  - 077718/WT_/Wellcome Trust/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20100722
PL  - England
TA  - J Med Microbiol
JT  - Journal of medical microbiology
JID - 0224131
RN  - 0 (DNA Primers)
RN  - 0 (DNA, Bacterial)
RN  - EC 1.2.1.- (Glyceraldehyde-3-Phosphate Dehydrogenases)
SB  - IM
MH  - Animals
MH  - Bacteriological Techniques/*methods
MH  - Blood/*microbiology
MH  - Cats
MH  - DNA Primers/genetics
MH  - DNA, Bacterial/genetics/isolation & purification
MH  - Glyceraldehyde-3-Phosphate Dehydrogenases/genetics
MH  - HIV Infections/complications
MH  - Humans
MH  - Mass Screening/methods
MH  - Mycoplasma/genetics/*isolation & purification
MH  - Mycoplasma Infections/*diagnosis
MH  - Polymerase Chain Reaction/*methods/standards
MH  - Reference Standards
MH  - Sensitivity and Specificity
MH  - South Africa
MH  - United Kingdom
PMC - PMC3090618
EDAT- 2010/07/24 06:00
MHDA- 2010/11/05 06:00
PMCR- 2010/11/01
CRDT- 2010/07/24 06:00
PHST- 2010/07/24 06:00 [entrez]
PHST- 2010/07/24 06:00 [pubmed]
PHST- 2010/11/05 06:00 [medline]
PHST- 2010/11/01 00:00 [pmc-release]
AID - 1285 [pii]
AID - 10.1099/jmm.0.021691-0 [doi]
PST - ppublish
SO  - J Med Microbiol. 2010 Nov;59(Pt 11):1285-1292. doi: 10.1099/jmm.0.021691-0. Epub 
      2010 Jul 22.