We evaluated a lyophilized CRISPR-Cas12 assay for SARS-CoV-2 detection (Lyo-CRISPR SARS-CoV-2 kit) based on reverse transcription, isothermal amplification, and CRISPR-Cas12 reaction. From a total of 210 RNA samples extracted from nasopharyngeal swabs using spin columns, the Lyo-CRISPR SARS-CoV-2 kit detected 105/105 (100%; 95% confidence interval (CI): 96.55–100) positive samples and 104/105 (99.05%; 95% CI: 94.81–99.97) negative samples that were previously tested using commercial RT-qPCR. The estimated overall Kappa index was 0.991, reflecting an almost perfect concordance level between the two diagnostic tests. An initial validation test was also performed on 30 nasopharyngeal samples collected in lysis buffer, in which the Lyo-CRISPR SARS-CoV-2 kit detected 20/21 (95.24%; 95% CI: 76.18–99.88) positive samples and 9/9 (100%; 95% CI: 66.37–100) negative samples. The estimated Kappa index was 0.923, indicating a strong concordance between the test procedures. The Lyo-CRISPR SARS-CoV-2 kit was suitable for detecting a wide range of RT-qPCR-positive samples (cycle threshold range: 11.45–36.90) and dilutions of heat-inactivated virus (range: 2.5–100 copies/µL); no cross-reaction was observed with the other respiratory pathogens tested. We demonstrated that the performance of the Lyo-CRISPR SARS-CoV-2 kit was similar to that of commercial RT-qPCR, as the former was highly sensitive and specific, timesaving (1.5 h), inexpensive, and did not require sophisticated equipment. The use of this kit would reduce the time taken for diagnosis and facilitate molecular diagnosis in low-resource laboratories.
Misiones province covers < 1% of Argentina’s land area yet harbors > 50% of the country’s biodiversity, with a significant remnant of Atlantic Forest, a global biodiversity hotspot. Approximately 540,000 ha of this native forest is protected, with the remaining areas facing threats from ongoing land conversion, an expanding road network, and a growing rural population. A prior study incorporated noninvasive data on five carnivores into a multifaceted cost analysis to define the optimal location for a multispecies biological corridor, with the goal of enhancing landscape connectivity among protected areas. Subsequent analyses, with an updated framework, emphasized management strategies that balanced human–wildlife coexistence and habitat needs. Building on these efforts, our study applied ecological niche modeling to data located by conservation detection dogs, with genetics used to confirm species identity, and two land-use scenarios, to predict potential distributions of three game species—lowland tapir (Tapirus terrestris), white-lipped peccary (Tayassu pecari), and collared peccary (Pecari tajacu)—that are not only threatened by poaching, road mortality, and habitat loss but also serve as essential prey for carnivores. We assessed the suitability of unique and overlapping vegetation types, within and outside of protected areas, as well as within this multispecies corridor, identifying zones of high conservation concern that underscore the need for integrated planning of predators and prey. These results highlight that ensuring the long-term viability of wildlife across the heterogeneous land-use matrices of Misiones requires going beyond protected areas to promote functional connectivity, restore degraded habitats, and balance human–wildlife needs.
About Me
Geneticist. PhD student at Buenos Aires University. Graduated from Misiones University (UNaM), Argentina.
Geneticist. PhD student at Buenos Aires University. Graduated from Misiones University (UNaM), Argentina.