{"doi":"10.3390/pathogens13010062","title":"Heat Inactivation of Nipah Virus for Downstream Single-Cell RNA Sequencing Does Not Interfere with Sample Quality","abstract":"Single-cell RNA sequencing (scRNA-seq) technologies are instrumental to improving our understanding of virus-host interactions in cell culture infection studies and complex biological systems because they allow separating the transcriptional signatures of infected versus non-infected bystander cells. A drawback of using biosafety level (BSL) 4 pathogens is that protocols are typically developed without consideration of virus inactivation during the procedure. To ensure complete inactivation of virus-containing samples for downstream analyses, an adaptation of the workflow is needed. Focusing on a commercially available microfluidic partitioning scRNA-seq platform to prepare samples for scRNA-seq, we tested various chemical and physical components of the platform for their ability to inactivate Nipah virus (NiV), a BSL-4 pathogen that belongs to the group of nonsegmented negative-sense RNA viruses. The only step of the standard protocol that led to NiV inactivation was a 5 min incubation at 85 °C. To comply with the more stringent biosafety requirements for BSL-4-derived samples, we included an additional heat step after cDNA synthesis. This step alone was sufficient to inactivate NiV-containing samples, adding to the necessary inactivation redundancy. Importantly, the additional heat step did not affect sample quality or downstream scRNA-seq results.","journal":"Pathogens","year":2024,"id":441753,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":8,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9589,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2024-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":232293,"name":"Judith Olejnik","orcid":"0000-0002-1748-7981","position":1,"is_corresponding":false},{"id":337525,"name":"Mitchell R. White","orcid":"0000-0001-6314-9353","position":2,"is_corresponding":false},{"id":232288,"name":"Jessie Huang","orcid":"0000-0002-5297-3563","position":3,"is_corresponding":false},{"id":669756,"name":"Jacquelyn Turcinovic","orcid":"0000-0002-7669-223X","position":4,"is_corresponding":false},{"id":821412,"name":"Baylee Heiden","orcid":"0000-0001-7087-6997","position":5,"is_corresponding":false},{"id":260367,"name":"Pushpinder Bawa","orcid":"0000-0002-8370-3638","position":6,"is_corresponding":false},{"id":551613,"name":"Christopher J. Williams","orcid":"0000-0002-5808-8768","position":7,"is_corresponding":false},{"id":1255299,"name":"Nickolas G. Gorham","orcid":null,"position":8,"is_corresponding":false},{"id":397494,"name":"Yuriy O. Alekseyev","orcid":"0000-0001-6105-8861","position":9,"is_corresponding":false},{"id":266494,"name":"John H. Connor","orcid":"0000-0002-8867-7256","position":10,"is_corresponding":false},{"id":228731,"name":"Darrell N. Kotton","orcid":"0000-0002-9604-8476","position":11,"is_corresponding":false},{"id":232302,"name":"Elke Mühlberger","orcid":"0000-0003-3547-9376","position":12,"is_corresponding":false},{"id":232289,"name":"Adam J. Hume","orcid":"0000-0001-8454-3472","position":0,"is_corresponding":true}],"reference_count":49,"raw_metadata":null,"created_at":"2026-07-19T02:01:11.152920Z","pmid":"38251369","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}