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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.

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Related in: MedlinePlus

Amplification curves from multiplex real-time PCR assays.Figure 1A. Real-time PCR amplification curves for body lice using a partial Phum_PHUM540560 gene in the FAM channel (495–520). Figure 1B. Amplification curves for head licee louse using a partial Phum_PHUM540560 gene in the VIC channel (522–544).
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pone-0058088-g002: Amplification curves from multiplex real-time PCR assays.Figure 1A. Real-time PCR amplification curves for body lice using a partial Phum_PHUM540560 gene in the FAM channel (495–520). Figure 1B. Amplification curves for head licee louse using a partial Phum_PHUM540560 gene in the VIC channel (522–544).

Mentions: The multiplex real-time PCR assay clearly identified and simultaneously differentiated among the 142 lice included in this work. Specifically, the signal emitted by the FAM-labeled probe was detected only in the body louse samples, whereas the signal emitted by the VIC-labeled probe was detected only in the head louse samples (Figure 2). No signals were detected in the non-template controls (NTCs). The Ct values obtained in this assay are outlined in Table S1. The sequencing of the 142 PCR products has confirmed our results. In addition, 100% of the samples that were positive for the FAM-labeled probe contained sequences specific to body lice, and 100% of the samples that were positive for the VIC-labeled probe contained sequences specific to head lice (100% sensitivity and 100% specificity; data not shown).


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)

Amplification curves from multiplex real-time PCR assays.Figure 1A. Real-time PCR amplification curves for body lice using a partial Phum_PHUM540560 gene in the FAM channel (495–520). Figure 1B. Amplification curves for head licee louse using a partial Phum_PHUM540560 gene in the VIC channel (522–544).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0058088-g002: Amplification curves from multiplex real-time PCR assays.Figure 1A. Real-time PCR amplification curves for body lice using a partial Phum_PHUM540560 gene in the FAM channel (495–520). Figure 1B. Amplification curves for head licee louse using a partial Phum_PHUM540560 gene in the VIC channel (522–544).
Mentions: The multiplex real-time PCR assay clearly identified and simultaneously differentiated among the 142 lice included in this work. Specifically, the signal emitted by the FAM-labeled probe was detected only in the body louse samples, whereas the signal emitted by the VIC-labeled probe was detected only in the head louse samples (Figure 2). No signals were detected in the non-template controls (NTCs). The Ct values obtained in this assay are outlined in Table S1. The sequencing of the 142 PCR products has confirmed our results. In addition, 100% of the samples that were positive for the FAM-labeled probe contained sequences specific to body lice, and 100% of the samples that were positive for the VIC-labeled probe contained sequences specific to head lice (100% sensitivity and 100% specificity; data not shown).

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