Evidence map›Paper›PMID 31924251›Full record

ArticleParasites & vectors2020

Highly specific PCR-RFLP assays for karyotyping the widespread 2Rb inversion in malaria vectors of the Anopheles gambiae complex.

Raquel Montanez-Gonzalez, Verena Pichler, Maria Calzetta, Rachel R Love, Alexandra Vallera, Lydia Schaecher, Beniamino Caputo, Marco Pombi, Vincenzo Petrarca, Alessandra Della Torre and 1 more

Open access · goldAbstract read
In one paragraph

Article in Parasites & vectors, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

0numbers the graph read from it
0cells of the map it votes in
6citing papers in PubMed
2.2field-weighted citation impact, top 12% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

6 citing papers in PubMed, 15 citations in OpenAlex.

  1. Article
  2. Review
  3. Article
  4. Article
  5. Article
  6. Inversion Genotyping in theG3 (Bethesda, Md.) · 2020
    Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

11 authors at 2 institutions in 2 countries.

Raquel Montanez-GonzalezEck Institute for Global Health, & Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, 46556, USA.
Verena PichlerDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy.
Maria CalzettaDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy.
Rachel R LoveEck Institute for Global Health, & Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, 46556, USA.
Alexandra ValleraEck Institute for Global Health, & Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, 46556, USA.
Lydia SchaecherEck Institute for Global Health, & Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, 46556, USA.
Beniamino CaputoDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy.
Marco PombiDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy.
Vincenzo PetrarcaDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy.
Alessandra Della TorreDipartimento di Sanità Pubblica e Malattie Infettive, Università "La Sapienza", Istituto Pasteur-Fondazione Cenci-Bolognetti, 00185, Rome, Italy. alessandra.dellatorre@uniroma1.it.
Nora J BesanskyEck Institute for Global Health, & Department of Biological Sciences, University of Notre Dame, Notre Dame, IN, 46556, USA. nbesansk@nd.edu.ORCID http://orcid.org/0000-0003-0646-0721
Istituto Pasteur · ITUniversity of Notre Dame · US

Funding

Empowering functional genomics of An. gambiae through inversion genotypingR01AI125360 · NIAID · UNIVERSITY OF NOTRE DAME · PI BESANSKY, NORA JESSIE · 2017 to 2021
$2.5M
Bill and Melinda Gates Foundation Target MalariaNIAID NIH HHS R01 AI125360Sapienza Università di Roma Progetti di Ricerca Università SAPIENZA 2018Silicon Valley Community Foundation Open Philanthropy Project FundUniversity of Notre Dame Glynn Family Honors ProgramUniversity of Notre Dame Kinesis-Fernández Richards Family Fellowship
6 · The paper itself

Abstract

backgroundChromosomal inversion polymorphisms play a role in adaptation to heterogeneous environments. Inversion polymorphisms are implicated in the very high ecological flexibility of the three main malaria vector species of the Afrotropical Anopheles gambiae complex, facilitating the exploitation of anthropogenic environmental modifications and promoting a strong association with humans. In addition to extending the species' spatial and temporal distribution, inversions are associated with epidemiologically relevant mosquito behavior and physiology, underscoring their medical importance. We here present novel PCR-RFLP based assays strongly predictive of genotype for the cosmopolitan 2Rb inversion in An. coluzzii and An. gambiae, a development which overcomes the numerous constraints inherent to traditional cytological karyotyping.

methodsWe designed PCR-RFLP genotyping assays based on tag SNPs previously computationally identified as strongly predictive (> 95%) of 2Rb genotype. We targeted those tags whose alternative allelic states destroyed or created the recognition site of a commercially available restriction enzyme, and designed assays with distinctive cleavage profiles for each inversion genotype. The assays were validated on 251 An. coluzzii and 451 An. gambiae cytologically karyotyped specimens from nine countries across Africa and one An. coluzzii laboratory colony.

resultsFor three tag SNPs, PCR-RFLP assays (denoted DraIII, MspAI, and TatI) reliably produced robust amplicons and clearly distinguishable electrophoretic profiles for all three inversion genotypes. Results obtained with the DraIII assay are ≥ 95% concordant with cytogenetic assignments in both species, while MspAI and TatI assays produce patterns highly concordant with cytogenetic assignments only in An. coluzzii or An. gambiae, respectively. Joint application of species-appropriate pairs of assays increased the concordance levels to > 99% in An. coluzzii and 98% in An. gambiae. Potential sources of discordance (e.g. imperfect association between tag and inversion, allelic dropout, additional polymorphisms in the restriction target site, incomplete or failed restriction digestion) are discussed.

conclusionsThe availability of highly specific, cost effective and accessible molecular assays for genotyping 2Rb in An. gambiae and An. coluzzii allows karyotyping of both sexes and all developmental stages. These novel tools will accelerate deeper investigations into the role of this ecologically and epidemiologically important chromosomal inversion in vector biology.

Indexed as

AfricaAnimalsAnophelesChromosome InversionCytogeneticsHumansKaryotypingMalariaMosquito VectorsPolymerase Chain ReactionPolymorphism, GeneticPolymorphism, Restriction Fragment LengthAnopheles gambiae complexChromosomal inversionInversion genotypingMalaria vectorMolecular karyotypingPCR-RFLPTag SNP

Identifiers

PMID31924251
PMCPMC6954590
OpenAlexW3005861671

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.