ArticleTuberculosis (Edinburgh, Scotland)2021
Using genomic DNA copies to enumerate Mycobacterium tuberculosis load in macaque tissue samples.
Article in Tuberculosis (Edinburgh, Scotland), 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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Who cites it
4 citing papers in PubMed, 4 citations in OpenAlex.
- Application and prospects of veterinary diagnostic techniques in laboratory monkeys.Frontiers in veterinary science · 2026Review
- CountFU: Ditching CFUs for qPCR enumeration of Mycobacterium tuberculosis and a call to join the revolution.Tuberculosis (Edinburgh, Scotland) · 2025Article
- Quantification ofWorld journal of hepatology · 2025Article
- Advancing tuberculosis diagnosis and management in cynomolgus macaques using Xpert MTB/RIF ultra assay.Scientific reports · 2024Article
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
Abstract
It is important to accurately quantify Mycobacterium tuberculosis (Mtb) load in laboratory-based tuberculosis (TB) research. This study aims to determine if real-time quantitative PCR (qPCR) and digital PCR (dPCR) can be used instead of colony forming unit (CFU) enumeration, to quantify Mtb load in rhesus and cynomolgus macaque tissue samples. Tissue samples of actively infected high Mtb-burden rhesus and cynomolgus macaques were selected from historic sample collections. CFUs were enumerated by plating, and Chelex-extracted genomic DNA used to quantify bacterial load by qPCR and dPCR. Three genes, sigA, 16S and CFP10, were assessed for their ability to quantify Mtb. All genes showed comparable quantification of Mtb between 2 and 20 000 copies/μl in the qPCR and 5-4000 copies/μl in the dPCR assay. The highest bacterial load was observed with dPCR, followed by qPCR, and CFU enumeration. Although the CFU count was consistently lower than the genomic copy numbers predicted by qPCR and dPCR, a significant correlation was observed. Quantification of Mtb by PCR was, however, only possible in higher-Mtb-load samples, suggesting that qPCR and dPCR quantification assays can predict bacterial load in actively infected and higher-Mtb-burden macaque tissue samples.
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Registered trials
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