An oligonucleotide-ligation assay (OLA) was developed and compared to a restriction fragment length polymorphism (RFLP) test for distinguishing between 294-bp polymerase chain reaction (PCR) amplification products of the 18S rRNA gene from Cyclospora and Eimeria spp. The PCR/OLA correctly distinguished between three Cyclospora, three E. tenella, and one E. mitis strains and the ratio of positive to negative spectrophotometric absorbance (A490) values for each strain ranged from 4.086 to 15.280 (median 9.5). PCR/OLA provides a rapid, reliable, spectrophotometric alternative to PCR/RFLP.
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http://dx.doi.org/10.4315/0362-028x-62.6.682 | DOI Listing |
J Mol Diagn
January 2025
Department of Biomedical Engineering, Vanderbilt University, Nashville, Tennessee, United States. Electronic address:
Single nucleotide variations (SNVs) and polymorphisms (SNPs) are characteristic biomarkers in various biological contexts, including pathogen drug resistances and human diseases. Tools that lower the implementation barrier of molecular SNV detection methods would provide greater leverage of the expanding SNP/SNV database. The oligonucleotide ligation assay (OLA) is a highly specific means for detection of known SNVs and is especially powerful when coupled with polymerase chain reaction (PCR).
View Article and Find Full Text PDFViruses
July 2024
Center for Global Infectious Disease Research, Seattle Children's Research Institute, Seattle, WA 98109, USA.
The WHO currently recommends dolutegravir (DTG)-based ART for persons living with HIV infection in resource-limited-settings (RLS). To expand access to testing for HIV drug resistance (DR) to DTG in RLS, we developed probes for use in the oligonucleotide ligation assay (OLA)-Simple, a near-point of care HIV DR kit. Genotypic data from clinical trials and case reports were used to determine the mutations in HIV-1 integrase critical to identifying individuals with DTG-resistance at virologic failure of DTG-based ART.
View Article and Find Full Text PDFJ Clin Virol
May 2023
Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA; Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford University School of Medicine, Stanford, CA, USA. Electronic address:
Background: SARS-CoV-2 variant surveillance informs vaccine composition and decisions to de-authorize antibody therapies. Though detailed genetic characterization requires whole-genome sequencing, targeted mutation analysis may complement pandemic surveillance efforts.
Methods: This study investigated the qualitative performance of a multiplex oligonucleotide ligation assay targeting 19 spike mutations using 192 whole genome sequenced upper respiratory samples representing SARS-CoV-2 variants of concern.
Med Trop Sante Int
December 2022
Faculté des sciences, Laboratoire de biologie et physiologie des organismes animaux, Université de Douala, B.P. 24 157 Douala, Cameroun.
Background: The effectiveness of Long-Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) in controlling malaria vectors is hampered by the resistance of anopheles to insecticides. A good knowledge of the breeding sites and of the resistance profile of the vectors could facilitate the development of an appropriate control strategy. This study looks at the larval ecology and the susceptibility profile of s.
View Article and Find Full Text PDFObjectives: Assess the impact of pre-treatment high-frequency and low-frequency drug-resistant HIV variants on long-term outcomes of first-line efavirenz-based antiretroviral therapy (ART).
Design: Prospective observational study.
Methods: Participants' pre-treatment plasma RNA had two sections of HIV pol encoding reverse transcriptase sequenced (Illumina, MiSeq) using unique molecular identifiers to detect wild-type (pre-treatment drug-resistant variants less than 1% of viral quasispecies), low-frequency (1-9%) or high-frequency drug-resistant variants (10-100%).
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