Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites DOI Creative Commons
Sandrine Musa,

Theo Hemberle,

Staffan Bensch

et al.

Frontiers in Cellular and Infection Microbiology, Journal Year: 2024, Volume and Issue: 14

Published: April 29, 2024

Avian haemosporidian parasites are useful model organisms to study the ecology and evolution of parasite-host interactions due their global distribution extensive biodiversity. Detection these has evolved from microscopic examination PCR-based methods, with mitochondrial cytochrome b gene serving as barcoding region. However, standard PCR protocols used for screening identification purposes have limitations in detecting mixed infections generating phylogenetically informative data short amplicon lengths. To address issues, we developed a novel genus-specific nested protocol targeting avian parasites. The underwent rigorous testing utilizing large dataset comprising blood samples Malagasy birds three distinct Passeriformes families. Furthermore, validation was done by examining smaller datasets two other laboratories employing divergent master mixes different bird species. Comparative analyses were conducted between outcomes those obtained through widely method. enables specific Plasmodium , Haemoproteus ( Parahaemoproteus ), Leucocytozoon demonstrated comparable sensitivity notable improvements infections. In addition, phylogenetic resolution is improved amplification longer fragments, leading better understanding biodiversity evolution. Overall, represents valuable addition detection methodologies, facilitating comprehensive studies on parasite ecology, epidemiology,

Language: Английский

Raising the bar: genus-specific nested PCR improves detection and lineage identification of avian haemosporidian parasites DOI Creative Commons
Sandrine Musa,

Theo Hemberle,

Staffan Bensch

et al.

Frontiers in Cellular and Infection Microbiology, Journal Year: 2024, Volume and Issue: 14

Published: April 29, 2024

Avian haemosporidian parasites are useful model organisms to study the ecology and evolution of parasite-host interactions due their global distribution extensive biodiversity. Detection these has evolved from microscopic examination PCR-based methods, with mitochondrial cytochrome b gene serving as barcoding region. However, standard PCR protocols used for screening identification purposes have limitations in detecting mixed infections generating phylogenetically informative data short amplicon lengths. To address issues, we developed a novel genus-specific nested protocol targeting avian parasites. The underwent rigorous testing utilizing large dataset comprising blood samples Malagasy birds three distinct Passeriformes families. Furthermore, validation was done by examining smaller datasets two other laboratories employing divergent master mixes different bird species. Comparative analyses were conducted between outcomes those obtained through widely method. enables specific Plasmodium , Haemoproteus ( Parahaemoproteus ), Leucocytozoon demonstrated comparable sensitivity notable improvements infections. In addition, phylogenetic resolution is improved amplification longer fragments, leading better understanding biodiversity evolution. Overall, represents valuable addition detection methodologies, facilitating comprehensive studies on parasite ecology, epidemiology,

Language: Английский

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