Maximizing viral nucleic acid yield from passive samplers: Evaluating elution and extraction protocols.

The COVID-19 pandemic has underscored the need for effective viral tracking in aqueous environments, particularly for non-enteric viruses. Despite advances in wastewater monitoring, surveillance of viruses in freshwater remains limited due to traditional sampling challenges. This study refines GAC-based passive sampling protocols by determining optimal extraction and elution methods for enhancing the recovery of viral nucleic acids in freshwater.

Enhanced detection of viruses for improved water safety.

Human viruses pose a significant health risk in freshwater environments, but current monitoring methods are inadequate for detecting viral presence efficiently. We evaluated a novel passive in-situ concentration method using granular activated carbon (GAC). This study detected and quantified eight enteric and non-enteric, pathogenic viruses in a freshwater recreational lake in paired grab and GAC passive samples.

Simultaneous detection of SARS-CoV-2, influenza A, respiratory syncytial virus, and measles in wastewater by multiplex RT-qPCR.

A multiplex quantitative reverse transcription polymerase chain reaction (RT-qPCR)-based method was designed for the simultaneous detection of influenza A, SARS-CoV-2, respiratory syncytial virus, and measles virus. The performance of the multiplex assay was compared to four monoplex assays for relative quantification using standard quantification curves. Results showed that the multiplex assay had comparable linearity and analytical sensitivity to the monoplex assays, and the quantification parameters of both assays demonstrated minimal differences.

Purification of the N- and C-terminal subdomains of recombinant heavy chain fragment C of botulinum neurotoxin serotype C.

The N-terminal and C-terminal portions of the heavy chain fragment C from botulinum neurotoxin serotype C [rBoNT(HC)] were expressed in Pichia pastoris and purified by ion-exchange chromotography (IEC). The N-terminal fragment, rBoNTC(Hc)-N, was purified in three IEC steps: a Q Sepharose Fast Flow (FF) capture step followed by a negative SP Sepharose FF step, and finally, Q Sepharose FF as a polishing step. The purification process resulted in greater than 90% pure rBoNTC(Hc)-N based on SDS-PAGE, and yielded up to 1.

Cell bank characterization and fermentation optimization for production of recombinant heavy chain C-terminal fragment of botulinum neurotoxin serotype E (rBoNTE(H(c)): antigen E) by Pichia pastoris.

A process was developed for production of a candidate vaccine antigen, recombinant C-terminal heavy chain fragment of the botulinum neurotoxin serotype E, rBoNTE(H(c)) in Pichia pastoris. P. pastoris strain GS115 was transformed with the rBoNTE(H(c)) gene inserted into pHILD4 Escherichia coli-P.