Biology subjects

Goyani, K. H.

Publications and source records attributed to Goyani, K. H..

3 recordsLinked to original sources

bioRxiv2023

Estimating limit of detection of M. tuberculosis DNA as an analyte released from a chemically coated solid matrix and using a WHO-approved CB-NAAT platform.

The limit of detection of Mtb (Mycobacterium tuberculosis) bacilli was evaluated by detection of released DNA from a chemically treated cellulose matrix. Loopful of the Mtb cells were suspended in phosphate buffer (pH 6.8), DNA extracted from it and quantified using a commercial Mtb real time PCR quantitation kit. Synthetic sputum, prepared using acrylamides as the matrix of choice was mixed with defined number of Mtb bacilli and spotted onto the cellulose matrix. Mtb DNA from the matrix was released into a DNA-release buffer and detected using a WHO-approved CB-NAAT detection platform. One hundred and fifty, 500 and 1000 but not 10 Mtb bacilli/mL of synthetic sputum could be detected by this protocol. The result underlined the potential of cellulose matrix as a potent, room temperature, Mtb-infected sputum transportation, storage and DNA-release device for sensitive detection of Mtb using Mtb Xpert Ultra CB-NAAT as the WHO-approved Mtb test of choice.

molecular biology↗

bioRxiv2023

Sputum-spotted solid matrix designed to release diagnostic-grade Mycobacterium tuberculosis DNA demonstrate optimal biocontainment property

Biosafety quotient of a reagent-coated cellulose matrix designed to release Mycobacterium tuberculosis (Mtb) DNA from sputum deposited onto it, was determined. Thirty-seven sputum samples infected with Mtb and 5 sputum samples from healthy individuals were processed and spotted onto the TBSend cards. Live Mtb bacilli were attempted for rescue from the spotted TBSend cards by washing them in phosphate buffer under mild shaking conditions. No live Mtb bacilli could be detected by the BACTEC MGIT 960 TB System from both Mtb infected as well as non-infected, sample-spotted TBSend cards when they were washed at two different time-points, viz., 1 minute and 15 hours from the time of spotting of the cards with sputum mixed with the preprocessing "spotting" buffer. The study reiterated published finding that chaotropic agents, which is an active component of the TBSend card module, have cent per cent bactericidal properties with regard to Mtb.

microbiology↗

bioRxiv2021

Mutations within the Open Reading Frame (ORF) including Ochre stop codon of the Surface Glycoprotein gene of SARS-CoV-2 virus erase potential seed location motifs of human non-coding microRNAs.

MicroRNA are short and non-coding RNA, 18-25 nucleotides in length. They are produced at the early stage of viral infection. The roles played by cellular miRNAs and miRNA-mediated gene-silencing in the COVID-19 epidemic period is critical in order to develop novel therapeutics. We analysed SARS-CoV-2 Surface Glycoprotein (S) nucleotide sequence originating from India as well as Iran, Australia, Germany, Italy, Russia, China, Japan and Turkey and identified mutation in potential seed location of several human miRNA. Seventy single nucleotide polymorphisms (SNP) were detected in the S gene out of which, 36, 32 and 2 were cases of transitions, transversions and deletions respectively. Eleven human miRNA targets were identified on the reference S gene sequence with a score >80 in the miRDB database. Mutation A845S erased a common binding site of 7 human miRNA (miR-195-5p, miR-16-5p, miR-15b-5p, miR-15a-5p, miR-497-5p, miR-424-5p and miR-6838-5p). A synonymous mutation altered the wild type Ochre stop codon within the S gene sequence (Italy) to Opal thereby changing the seed sequence of miR-511-3p. Similar (synonymous) mutations were detected at amino acid position 659 and 1116 of the S gene where amino acids serine and threonine were retained, abolishing potential seed location for miR-219a-1-3p and miR-20b-3p respectively. The significance of this finding in reference to the strategy to use synthetic miRNA combinations as a novel therapeutic tool is discussed.

molecular biology↗