Primers and visualization of LAMP: A rapid molecular identification method for Liposcelis entomophila (Enderlein) (Psocodea: Liposcelididae)

Estimation of postmortem interval (PMI) is paramount in modern forensic investigation. After the disappearance of the early postmortem phenomena conventionally used to estimate PMI, entomologic evidence provides important indicators for PMI estimation. The age of the oldest fly larvae or pupae can be estimated to pinpoint the time of oviposition, which is considered the minimum PMI (PMImin). The development rate of insects is usually temperature dependent and species specific. Therefore, species identification is mandatory for PMImin estimation using entomological evidence. The classical morphological identification method cannot be applied when specimens are damaged or have not yet matured. To overcome this limitation, some investigators employ molecular identification using mitochondrial cytochrome c oxidase subunit I (COI) nucleotide sequences. The molecular identification method commonly uses Sanger’s nucleotide sequencing and molecular phylogeny, which are complex and time consuming and constitute another obstacle for forensic investigators. In this study, instead of using conventional Sanger’s nucleotide sequencing, single-nucleotide polymorphisms (SNPs) in the COI gene region, which are unique between fly species, were selected and targeted for single-base extension (SBE) technology. These SNPs were genotyped using a SNaPshot® kit. Eleven Calliphoridae and seven Sarcophagidae species were covered. To validate this genotyping, fly DNA samples (103 adults, 84 larvae, and 4 pupae) previously confirmed by DNA barcoding were used. This method worked quickly with minimal DNA, providing a potential alternative to conventional DNA barcoding. Consisting of only a few simple electropherogram peaks, the results were more straightforward compared with those of the conventional DNA barcoding produced by Sanger’s nucleotide sequencing.

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Development of Molecular Identification Method for Genus Alexandrium (Dinophyceae) Using Whole-Cell FISH

Marine Biotechnology ◽

10.1007/s10126-004-0424-2 ◽

2005 ◽

Vol 7 (3) ◽

pp. 215-222 ◽

Cited By ~ 12

Author(s):

Choong-Jae Kim ◽

Chang-Hoon Kim ◽

Yoshihiko Sako

Keyword(s):

Molecular Identification ◽

Whole Cell ◽

Identification Method

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Validation of an Effective Protocol for Culicoides Latreille (Diptera: Ceratopogonidae) Detection Using eDNA Metabarcoding

Insects ◽

10.3390/insects12050401 ◽

2021 ◽

Vol 12 (5) ◽

pp. 401

Author(s):

Yoamel Milián-García ◽

Lauren A. A. Janke ◽

Robert G. Young ◽

Aruna Ambagala ◽

Robert H. Hanner

Keyword(s):

Molecular Identification ◽

Molecular Detection ◽

Morphological Characteristics ◽

Species Level ◽

Morphological Identification ◽

Culicoides Species ◽

Nacl Solution ◽

Identification Method

eDNA metabarcoding is an effective molecular-based identification method for the biosurveillance of flighted insects. An eDNA surveillance approach maintains specimens for secondary morphological identification useful for regulatory applications. This study identified Culicoides species using eDNA metabarcoding and compared these results to morphological identifications of trapped specimens. Insects were collected using ultraviolet (UV) lighted fan traps containing a saturated salt (NaCl) solution from two locations in Guelph, Ontario, Canada. There were forty-two Culicoides specimens collected in total. Molecular identification detected four species, C. biguttatus, C. stellifer, C. obsoletus, and C. mulrennani. Using morphological identification, two out of these four taxonomic ranks were confirmed at the species level (C. biguttatus and C. stellifer) and one was confirmed at the subgenus level (Avaritia [C. obsoletus]). No molecular detection of Culicoides species occurred in traps with an abundance of less than three individuals per taxon. The inconsistency in identifying Culicoides specimens to the species level punctuates the need for curated DNA reference libraries for Culicoides. In conclusion, the saturated salt (NaCl) solution preserved the Culicoides’ morphological characteristics and the eDNA.

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Sponge-Cell Culture? A Molecular Identification Method for Sponge Cells

Marine Biotechnology ◽

10.1007/s10126-002-0090-1 ◽

2003 ◽

Vol 5 (5) ◽

pp. 443-449 ◽

Cited By ~ 13

Author(s):

Detmer Sipkema ◽

Hans G.H.J. Heilig ◽

Antoon D.L. Akkermans ◽

Ronald Osinga ◽

Johannes Onji ◽

...

Keyword(s):

Cell Culture ◽

Molecular Identification ◽

Sponge Cell ◽

Identification Method

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Electron microscopy in diagnostic virology

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100110702 ◽

1984 ◽

Vol 42 ◽

pp. 70-73

Author(s):

S. E. Miller

Keyword(s):

Electron Microscopy ◽

Tissue Culture ◽

Molecular Identification ◽

Point Of View ◽

Negative Stain ◽

Viral Diagnosis ◽

Electron Microscopist ◽

Diagnostic Virology

The techniques for detecting viruses are many and varied including FAT, ELISA, SPIRA, RPHA, SRH, TIA, ID, IEOP, GC (1); CF, CIE (2); Tzanck (3); EM, IEM (4); and molecular identification (5). This paper will deal with viral diagnosis by electron microscopy and will be organized from the point of view of the electron microscopist who is asked to look for an unknown agent--a consideration of the specimen and possible agents rather than from a virologist's view of comparing all the different viruses. The first step is to ascertain the specimen source and select the method of preparation, e. g. negative stain or embedment, and whether the sample should be precleared by centrifugation, concentrated, or inoculated into tissue culture. Also, knowing the type of specimen and patient symptoms will lend suggestions of possible agents and eliminate some viruses, e. g. Rotavirus will not be seen in brain, nor Rabies in stool, but preconceived notions should not prejudice the observer into missing an unlikely pathogen.

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