Investigation of human haemotropic Mycoplasma infections using a novel generic haemoplasma qPCR assay on blood samples and blood smears.

Journal of Medical Microbiology
Séverine TaskerH Neimark

Abstract

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 ...Continue Reading

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Citations

Mar 6, 2013·New Zealand Veterinary Journal·E Barker, S Tasker
Oct 25, 2011·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Jane A SteerMike Hamon
Dec 23, 2011·Journal of Clinical Microbiology·Pálma SillóSarolta Kárpáti
May 16, 2012·BMC Veterinary Research·Emily N BarkerSéverine Tasker
Feb 23, 2013·The Journal of Veterinary Medical Science·Hinako SashidaRyô Harasawa
Dec 24, 2013·Vector Borne and Zoonotic Diseases·Sándor HornokRegina Hofmann-Lehmann
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Dec 15, 2010·Veterinary Microbiology·Emily N BarkerSéverine Tasker
Oct 20, 2015·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Rachel Hutchinson, Wendy Gibson
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Oct 30, 2020·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Elodie DesclouxDidier Raoult

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Datasets Mentioned

BETA
U95297

Methods Mentioned

BETA
PCR
electrophoresis
PCRs
light microscopy

Software Mentioned

Primer3Plus
iCycler iQ Optical System
DS Gene

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