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Relationship of taxonomic error to frequency of observation.

ABSTRACT: Biological nomenclature is the entry point to a wealth of information related to or associated with living entities. When applied accurately and consistently, communication between and among researchers and investigators is enhanced, leading to advancements in understanding and progress in research programs. Based on freshwater benthic macroinvertebrate taxonomic identifications, inter-laboratory comparisons of >900 samples taken from rivers, streams, and lakes across the U.S., including the Great Lakes, provided data on taxon-specific error rates. Using the error rates in combination with frequency of observation (FREQ; as a surrogate for rarity), six uncertainty/frequency classes (UFC) are proposed for approximately 1,000 taxa. The UFC, error rates, FREQ each are potentially useful for additional analyses related to interpreting biological assessment results and/or stressor response relationships, as weighting factors for various aspects of ecological condition or biodiversity analyses and helping set direction for taxonomic research and refining identification tools.

SUBMITTER: Stribling JB

PROVIDER: S-EPMC7660486 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature

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Relationship of taxonomic error to frequency of observation.

Stribling James B JB Leppo Erik W EW

PloS one 20201112 11

Biological nomenclature is the entry point to a wealth of information related to or associated with living entities. When applied accurately and consistently, communication between and among researchers and investigators is enhanced, leading to advancements in understanding and progress in research programs. Based on freshwater benthic macroinvertebrate taxonomic identifications, inter-laboratory comparisons of >900 samples taken from rivers, streams, and lakes across the U.S., including the Gre ...[more]

PMID: 33180842

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Project description:454 sequencing technology is a promising approach for characterizing HIV-1 populations and for identifying low frequency mutations. The utility of 454 technology for determining allele frequencies and linkage associations in HIV infected individuals has not been extensively investigated. We evaluated the performance of 454 sequencing for characterizing HIV populations with defined allele frequencies.We constructed two HIV-1 RT clones. Clone A was a wild type sequence. Clone B was identical to clone A except it contained 13 introduced drug resistant mutations. The clones were mixed at ratios ranging from 1% to 50% and were amplified by standard PCR conditions and by PCR conditions aimed at reducing PCR-based recombination. The products were sequenced using 454 pyrosequencing. Sequence analysis from standard PCR amplification revealed that 14% of all sequencing reads from a sample with a 50:50 mixture of wild type and mutant DNA were recombinants. The majority of the recombinants were the result of a single crossover event which can happen during PCR when the DNA polymerase terminates synthesis prematurely. The incompletely extended template then competes for primer sites in subsequent rounds of PCR. Although less often, a spectrum of other distinct crossover patterns was also detected. In addition, we observed point mutation errors ranging from 0.01% to 1.0% per base as well as indel (insertion and deletion) errors ranging from 0.02% to nearly 50%. The point errors (single nucleotide substitution errors) were mainly introduced during PCR while indels were the result of pyrosequencing. We then used new PCR conditions designed to reduce PCR-based recombination. Using these new conditions, the frequency of recombination was reduced 27-fold. The new conditions had no effect on point mutation errors. We found that 454 pyrosequencing was capable of identifying minority HIV-1 mutations at frequencies down to 0.1% at some nucleotide positions.Standard PCR amplification results in a high frequency of PCR-introduced recombination precluding its use for linkage analysis of HIV populations using 454 pyrosequencing. We designed a new PCR protocol that resulted in a much lower recombination frequency and provided a powerful technique for linkage analysis and haplotype determination in HIV-1 populations. Our analyses of 454 sequencing results also demonstrated that at some specific HIV-1 drug resistant sites, mutations can reliably be detected at frequencies down to 0.1%.

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Relationship of taxonomic error to frequency of observation.

Publications

Relationship of taxonomic error to frequency of observation.

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