Limits...
Distinguishing body lice from head lice by multiplex real-time PCR analysis of the Phum_PHUM540560 gene.

Drali R, Boutellis A, Raoult D, Rolain JM, Brouqui P - PLoS ONE (2013)

Bottom Line: The multiplex real-time PCR analysis enabled the body and head lice to be distinguished in two hours.This method is simple, with 100% specificity and sensitivity.We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.

View Article: PubMed Central - PubMed

Affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, Marseille, France.

ABSTRACT

Background: Body louse or head louse? Once removed from their environment, body and head lice are indistinguishable. Neither the morphological criteria used since the mid-18th century nor the various genetic studies conducted since the advent of molecular biology tools have allowed body lice and head lice to be differentiated. In this work, using a portion of the Phum_PHUM540560 gene from the body louse, we aimed to develop a multiplex real-time polymerase chain reaction (PCR) assay to differentiate between body and head lice in a single reaction.

Materials and methods: A total of 142 human lice were collected from mono-infested hosts from 13 countries on five continents. We first identified the louse clade using a cytochrome b (CYTB) PCR sequence alignment. We then aligned a fragment of the Phum_PHUM540560 gene amplified from head and body lice to design-specific TaqMan(©) FAM- and VIC-labeled probes.

Results: All the analyzed lice were Clade A lice. A total of 22 polymorphisms between the body and head lice were characterized. The multiplex real-time PCR analysis enabled the body and head lice to be distinguished in two hours. This method is simple, with 100% specificity and sensitivity.

Conclusions: We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.

Show MeSH

Related in: MedlinePlus

Primer and probe alignments with partial Phum_PHUM540560 gene sequences from body and head lice [35].A portion of the Phum_PHUM540560 gene sequences from body and head lice were aligned with the primers and probes designed for the multiplex RT-PCR assay. Part of the first exon spanings nucleotides 1 to 64 was analyzed. The forward and reverse primer sequences are boxed in black. The FAM- and VIC-labeled probe sequences are boxed in purple and green, respectively. The nucleotides in blue represent single-nucleotide polymorphisms that are specific to head lice. The nucleotides in black represent single-nucleotide polymorphisms that are specific to body lice. BL: body louse; HL: head louse; NW_002987859.1: Pediculus humanus corporis strain USDA 1103172108290 Phum_PHUM540560 (gene sequence available in GenBank).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3585238&req=5

pone-0058088-g001: Primer and probe alignments with partial Phum_PHUM540560 gene sequences from body and head lice [35].A portion of the Phum_PHUM540560 gene sequences from body and head lice were aligned with the primers and probes designed for the multiplex RT-PCR assay. Part of the first exon spanings nucleotides 1 to 64 was analyzed. The forward and reverse primer sequences are boxed in black. The FAM- and VIC-labeled probe sequences are boxed in purple and green, respectively. The nucleotides in blue represent single-nucleotide polymorphisms that are specific to head lice. The nucleotides in black represent single-nucleotide polymorphisms that are specific to body lice. BL: body louse; HL: head louse; NW_002987859.1: Pediculus humanus corporis strain USDA 1103172108290 Phum_PHUM540560 (gene sequence available in GenBank).

Mentions: The multiple alignments of the partial Phum_PHUM54056 gene sequences obtained from the six analyzed lice revealed 22 polymorphisms between body and head lice (Figure 1). The first exon contained two point mutations: a silent (CCA>CCC) transversion affecting codon 18 that would not change the amino acid and the I19N (ATT>AAT) mutation that would replace isoleucine with asparagine.


Distinguishing body lice from head lice by multiplex real-time PCR analysis of the Phum_PHUM540560 gene.

Drali R, Boutellis A, Raoult D, Rolain JM, Brouqui P - PLoS ONE (2013)

Primer and probe alignments with partial Phum_PHUM540560 gene sequences from body and head lice [35].A portion of the Phum_PHUM540560 gene sequences from body and head lice were aligned with the primers and probes designed for the multiplex RT-PCR assay. Part of the first exon spanings nucleotides 1 to 64 was analyzed. The forward and reverse primer sequences are boxed in black. The FAM- and VIC-labeled probe sequences are boxed in purple and green, respectively. The nucleotides in blue represent single-nucleotide polymorphisms that are specific to head lice. The nucleotides in black represent single-nucleotide polymorphisms that are specific to body lice. BL: body louse; HL: head louse; NW_002987859.1: Pediculus humanus corporis strain USDA 1103172108290 Phum_PHUM540560 (gene sequence available in GenBank).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585238&req=5

pone-0058088-g001: Primer and probe alignments with partial Phum_PHUM540560 gene sequences from body and head lice [35].A portion of the Phum_PHUM540560 gene sequences from body and head lice were aligned with the primers and probes designed for the multiplex RT-PCR assay. Part of the first exon spanings nucleotides 1 to 64 was analyzed. The forward and reverse primer sequences are boxed in black. The FAM- and VIC-labeled probe sequences are boxed in purple and green, respectively. The nucleotides in blue represent single-nucleotide polymorphisms that are specific to head lice. The nucleotides in black represent single-nucleotide polymorphisms that are specific to body lice. BL: body louse; HL: head louse; NW_002987859.1: Pediculus humanus corporis strain USDA 1103172108290 Phum_PHUM540560 (gene sequence available in GenBank).
Mentions: The multiple alignments of the partial Phum_PHUM54056 gene sequences obtained from the six analyzed lice revealed 22 polymorphisms between body and head lice (Figure 1). The first exon contained two point mutations: a silent (CCA>CCC) transversion affecting codon 18 that would not change the amino acid and the I19N (ATT>AAT) mutation that would replace isoleucine with asparagine.

Bottom Line: The multiplex real-time PCR analysis enabled the body and head lice to be distinguished in two hours.This method is simple, with 100% specificity and sensitivity.We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.

View Article: PubMed Central - PubMed

Affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, Marseille, France.

ABSTRACT

Background: Body louse or head louse? Once removed from their environment, body and head lice are indistinguishable. Neither the morphological criteria used since the mid-18th century nor the various genetic studies conducted since the advent of molecular biology tools have allowed body lice and head lice to be differentiated. In this work, using a portion of the Phum_PHUM540560 gene from the body louse, we aimed to develop a multiplex real-time polymerase chain reaction (PCR) assay to differentiate between body and head lice in a single reaction.

Materials and methods: A total of 142 human lice were collected from mono-infested hosts from 13 countries on five continents. We first identified the louse clade using a cytochrome b (CYTB) PCR sequence alignment. We then aligned a fragment of the Phum_PHUM540560 gene amplified from head and body lice to design-specific TaqMan(©) FAM- and VIC-labeled probes.

Results: All the analyzed lice were Clade A lice. A total of 22 polymorphisms between the body and head lice were characterized. The multiplex real-time PCR analysis enabled the body and head lice to be distinguished in two hours. This method is simple, with 100% specificity and sensitivity.

Conclusions: We confirmed that the Phum_PHUM540560 gene is a useful genetic marker for the study of lice.

Show MeSH
Related in: MedlinePlus