Background Body louse or head louse? Once removed from their environment, body and head lice are indistinguishable. 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. Introduction Body and head lice are hematophagous ectoparasites that are specific to humans [1] and have different ecologies. The body louse, and provided new perspectives for understanding the relationship between the biology and genetics of the louse [17]. More recently, a study comparing the transcriptional profiles of body and head lice reported that the two types of lice experienced a single, 752-base pair (bp) difference in the Phum_PHUM540560 gene, which encodes a hypothetical, 69-amino acids protein of unfamiliar function [18]. Based on the positioning of a portion of the two Phum_PHUM540560 gene sequences, we have designed a novel multiplex real-time PCR assay to efficiently differentiate, for the first time, between body and head lice collected from a mono-infested sponsor. This assay has been tested by analyzing a large collection of worldwide specimens belonging to Clade A, the only clade known to consist of both body and head lice. Materials and Methods Ethics statement Lice from foreign countries were from the private frozen collection of our laboratory (The URMITE/WHO Collaborative Study Center). The lice in that collection were required for numerous epidemiological and entomological studies or to perform diagnoses abroad and were sent to our laboratory like a WHO research facility. The specimens were collected according to the ethics laws of each country; however, because lice are not part of the human body, lice removed from individuals are not considered to be IFNW1 human samples in most countries. The body lice were collected from clothing, and the head lice were removed buy 21967-41-9 from hair, with the verbal consent of the infested individuals. Written consent was not obtainable in the majority of cases because most of the subjects were illiterate. However, in most instances, the investigator, local authorities and/or town chief authorized and were present when it was performed. The lice collected in France were from homeless individuals during a authorized epidemiological study (French Bioethics laws n 2011C814). Informed consent was from these subjects, and the study was authorized by the Comit de Safety des Personnes Sud Mediterrane I on January, 12, 2011 (ID RCB: 2010-A01406-33). The anonymity of the individuals who offered the lice used in the present genetic analysis was maintained. Sampling A total of 142 lice, including 88 body lice and 54 head lice, were collected from mono-infested human being hosts. The head lice were collected specifically from your hair, and the body lice were collected specifically from clothing. No lice were collected from your buy 21967-41-9 throat or the beard; the purpose of this precaution was the avoidance cross lice, as previously reported [7]. The strain info, geographic source and anatomical sources (body or head) of the analyzed lice are provided in Table 1. Table 1 The Clade A lice examined with this study and the results of the real-time PCR assay. DNA preparation Prior to DNA isolation, each louse was immersed in 70% ethanol for 15 min and was then rinsed twice in sterile water. Total genomic DNA was extracted using the QIAamp Cells Kit (QIAGEN, Hilden, Germany) according to the manufacturer’s instructions. The extracted DNA was assessed for amount and quality using a NanoDrop instrument (Thermo Scientific, Wilmington, United Kingdom) before becoming stored at ?20C [19]. Conventional PCR and sequencing Two standard PCR experiments were performed with buy 21967-41-9 this study. The 1st was performed to identify the Clades of the collected lice by amplifying and sequencing.