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2.
PLoS Negl Trop Dis ; 15(12): e0009787, 2021 12.
Article in English | MEDLINE | ID: mdl-34890398

ABSTRACT

Combined community health programs aiming at health education, preventive anti-parasitic chemotherapy, and vaccination of pigs have proven their potential to regionally reduce and even eliminate Taenia solium infections that are associated with a high risk of neurological disease through ingestion of T. solium eggs. Yet it remains challenging to target T. solium endemic regions precisely or to make exact diagnoses in individual patients. One major reason is that the widely available stool microscopy may identify Taenia ssp. eggs in stool samples as such, but fails to distinguish between invasive (T. solium) and less invasive Taenia (T. saginata, T. asiatica, and T. hydatigena) species. The identification of Taenia ssp. eggs in routine stool samples often prompts a time-consuming and frequently unsuccessful epidemiologic workup in remote villages far away from a diagnostic laboratory. Here we present "mail order" single egg RNA-sequencing, a new method allowing the identification of the exact Taenia ssp. based on a few eggs found in routine diagnostic stool samples. We provide first T. solium transcriptome data, which show extremely high mitochondrial DNA (mtDNA) transcript counts that can be used for subspecies classification. "Mail order" RNA-sequencing can be administered by health personnel equipped with basic laboratory tools such as a microscope, a Bunsen burner, and access to an international post office for shipment of samples to a next generation sequencing facility. Our suggested workflow combines traditional stool microscopy, RNA-extraction from single Taenia eggs with mitochondrial RNA-sequencing, followed by bioinformatic processing with a basic laptop computer. The workflow could help to better target preventive healthcare measures and improve diagnostic specificity in individual patients based on incidental findings of Taenia ssp. eggs in diagnostic laboratories with limited resources.


Subject(s)
Feces/parasitology , Sequence Analysis, RNA/methods , Taenia solium/genetics , Taeniasis/diagnosis , Taeniasis/parasitology , Animals , DNA, Mitochondrial/genetics , High-Throughput Nucleotide Sequencing , Humans , Ovum/chemistry , Postal Service , RNA, Helminth/genetics , Species Specificity , Taenia solium/classification , Taenia solium/isolation & purification
3.
Nat Biotechnol ; 39(8): 968-977, 2021 08.
Article in English | MEDLINE | ID: mdl-33875865

ABSTRACT

Several techniques are currently being developed for spatially resolved omics profiling, but each new method requires the setup of specific detection strategies or specialized instrumentation. Here we describe an imaging-free framework to localize high-throughput readouts within a tissue by cutting the sample into thin strips in a way that allows subsequent image reconstruction. We implemented this framework to transform a low-input RNA sequencing protocol into an imaging-free spatial transcriptomics technique (called STRP-seq) and validated it by profiling the spatial transcriptome of the mouse brain. We applied the technique to the brain of the Australian bearded dragon, Pogona vitticeps. Our results reveal the molecular anatomy of the telencephalon of this lizard, providing evidence for a marked regionalization of the reptilian pallium and subpallium. We expect that STRP-seq can be used to derive spatially resolved data from a range of other omics techniques.


Subject(s)
Gene Expression Profiling/methods , Molecular Imaging/methods , Tomography/methods , Algorithms , Animals , Brain/diagnostic imaging , Brain/metabolism , Brain Chemistry , Lizards , Mice , Transcriptome/genetics
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