Skip to main content Accessibility help

Identification of a novel PYP-1 gene in Sarcoptes scabiei and its potential as a serodiagnostic candidate by indirect-ELISA

  • Jing Xu (a1), Xing Huang (a2), Manli He (a1), Yongjun Ren (a3) (a4), Nengxing Shen (a1), Chunyan Li (a1), Ran He (a1), Yue Xie (a1), Xiaobin Gu (a1), Bo Jing (a1), Xuerong Peng (a5) and Guangyou Yang (a1)...


Scabies is a parasitic disease caused by the ectoparasite Sarcoptes scabiei, affecting different mammalian species, including rabbits, worldwide. In the present study, we cloned and expressed a novel inorganic pyrophosphatase, Ssc-PYP-1, from S. scabiei var. cuniculi. Immunofluorescence staining showed that native Ssc-PYP-1 was localized in the tegument around the mouthparts and the entire legs, as well as in the cuticle of the mites. Interestingly, obvious staining was also observed on the fecal pellets of mites and in the integument of the mites. Based on its good immunoreactivity, an indirect enzyme-linked immunosorbent assay (ELISA) using recombinant Ssc-PYP-1 (rSsc-PYP-1) as the capture antigen was developed to diagnose sarcoptic mange in naturally infected rabbits; the assay had a sensitivity of 92·0% and specificity of 93·6%. Finally, using the rSsc-PYP-1-ELISA, the Ssc-PYP-1 antibody from 10 experimentally infected rabbits could be detected from 1 week post-infection. This is the first report of S. scabiei inorganic pyrophosphatase and the protein could serve as a potential serodiagnostic candidate for sarcoptic mange in rabbits.


Corresponding author

Author for Correspondence: Guangyou Yang, E-mail:;


Hide All
Alasaad, S, Rossi, L, Heukelbach, J, Pérez, JM, Hamarsheh, O, Otiende, M and Zhu, XQ (2013) The neglected navigating web of the incomprehensibly emerging and re-emerging Sarcoptes mite. Infection, Genetics and Evolution 17, 253259.
Angelone-Alasaad, S, Min, AM, Pasquetti, M, Alagaili, AN, D'Amelio, S, Berrilli, F, Obanda, V, Gebely, MA, Soriguer, RC and Rossi, L (2015) Universal conventional and real-time PCR diagnosis tools for Sarcoptes scabiei. Parasites & Vectors 8, 587.
Arlian, LG, Morgan, MS, Rapp, CM and Vyszenski-Moher, DL (1996) The development of protective immunity in canine scabies. Veterinary Parasitology 62, 133142.
Bates, P (1999) Inter-and intra-specific variation within the genus Psoroptes (Acari: Psoroptidae). Veterinary Parasitology 83, 201217.
Beckham, SA, Boyd, SE, Reynolds, S, Willis, C, Johnstone, M, Mika, A, Simerská, P, Wijeyewickrema, LC, Smith, AI and Kemp, DJ (2009) Characterization of a serine protease homologous to house dust mite group 3 allergens from the scabies mite Sarcoptes scabiei. Journal of Biological Chemistry 284, 3441334422.
Bornstein, S, Zakrisson, G and Thebo, P (1994) Clinical picture and antibody response to experimental Sarcoptes scabiei var. Vulpes infection in red foxes (Vulpes vulpes). Acta Veterinaria Scandinavica 36, 509519.
Casais, R, Goyena, E, Martínez-Carrasco, C, de Ybáñez, RR, de Vega, FA, Ramis, G, Prieto, J and Berriatua, E (2013) Variable performance of a human derived Sarcoptes scabiei recombinant antigen ELISA in swine mange diagnosis. Veterinary Parasitology 197, 397403.
Casais, R, Dalton, KP, Millán, J, Balseiro, A, Oleaga, Á, Solano, P, Goyache, F, Prieto, JM and Parra, F (2014) Primary and secondary experimental infestation of rabbits (Oryctolagus cuniculus) with Sarcoptes scabiei from a wild rabbit: factors determining resistance to reinfestation. Veterinary Parasitology 203, 173183.
Casais, R, Millán, J, Rosell, JM, Dalton, KP and Prieto, JM (2015) Evaluation of an ELISA using recombinant Ssλ20ΔB3 antigen for the serological diagnosis of Sarcoptes scabiei infestation in domestic and wild rabbits. Veterinary Parasitology 214, 315321.
Chosidow, O (2000) Scabies and pediculosis. The Lancet 355, 819826.
Crowther, J and Walker, J (2009) The ELISA Guidebook, 2nd edn. Humana Press, New Jersey, US.
Engelman, D, Kiang, K, Chosidow, O, McCarthy, J, Fuller, C, Lammie, P, Hay, R, Steer, A and Members of the International Alliance for the Control of Scabies (IACS) (2013) Toward the global control of human scabies: introducing the international alliance for the control of scabies. PLoS Neglected Tropical Diseases 7, e2167.
Espiau, B, Lemercier, G, Ambit, A, Bringaud, F, Merlin, G, Baltz, T and Bakalara, N (2006) A soluble pyrophosphatase, a key enzyme for polyphosphate metabolism in Leishmania. Journal of Biological Chemistry 281, 15161523.
Fairchild, TA and Patejunas, G (1999) Cloning and expression profile of human inorganic pyrophosphatase. Biochimica et Biophysica Acta (BBA) – Gene Structure and Expression 1447, 133136.
Fukuyama, S, Nishimura, T, Yotsumoto, H, Gushi, A, Tsuji, M, Kanekura, T and Matsuyama, T (2010) Diagnostic usefulness of a nested polymerase chain reaction assay for detecting Sarcoptes scabiei DNA in skin scrapings from clinically suspected scabies. British Journal of Dermatology 163, 892894.
Galizzi, M, Bustamante, JM, Fang, J, Miranda, K, Soares Medeiros, LC, Tarleton, RL and Docampo, R (2013) Evidence for the role of vacuolar soluble pyrophosphatase and inorganic polyphosphate in Trypanosoma cruzi persistence. Molecular Microbiology 90, 699715.
Haas, N and Sterry, W (2001) The use of ELM to monitor the success of antiscabietic treatment. Archives of Dermatology 137, 16561657.
Hay, R, Steer, A, Engelman, D and Walton, S (2012) Scabies in the developing world—its prevalence, complications, and management. Clinical Microbiology and Infection 18, 313323.
Hay, RJ, Johns, NE, Williams, HC, Bolliger, IW, Dellavalle, RP, Margolis, DJ, Marks, R, Naldi, L, Weinstock, MA and Wulf, SK (2014) The global burden of skin disease in 2010: an analysis of the prevalence and impact of skin conditions. Journal of Investigative Dermatology 134, 15271534.
He, R, Shen, N, Lin, H, Gu, X, Lai, W, Peng, X and Yang, G (2017) Molecular characterization of calmodulin from Sarcoptes scabiei. Parasitology International 66, 16.
Heikinheimo, P, Tuominen, V, Ahonen, AK, Teplyakov, A, Cooperman, B, Baykov, A, Lahti, R and Goldman, A (2001) Toward a quantum-mechanical description of metal-assisted phosphoryl transfer in pyrophosphatase. Proceedings of the National Academy of Sciences of the United States of America 98, 31213126.
Heinonen, JK (2001) Biological Role of Inorganic Pyrophosphate. Springer, Boston, MA.
Heukelbach, J, Wilcke, T, Winter, B and Feldmeier, H (2005) Epidemiology and morbidity of scabies and pediculosis capitis in resource-poor communities in Brazil. British Journal of Dermatology 153, 150156.
Islam, MK, Miyoshi, T, Kasuga-Aoki, H, Isobe, T, Arakawa, T, Matsumoto, Y and Tsuji, N (2003) Inorganic pyrophosphatase in the roundworm Ascaris and its role in the development and molting process of the larval stage parasites. European Journal of Biochemistry 270, 28142826.
Islam, MK, Miyoshi, T, Yamada, M and Tsuji, N (2005) Pyrophosphatase of the roundworm Ascaris suum plays an essential role in the worm's molting and development. Infection and Immunity 73, 19952004.
Islam, MK, Miyoshi, T, Yamada, M, Alim, MA, Huang, X, Motobu, M and Tsuji, N (2006) Effect of piperazine (diethylenediamine) on the moulting, proteome expression and pyrophosphatase activity of Ascaris suum lung-stage larvae. Acta Tropica 99, 208217.
Jacobson, R (1998) Validation of serological assays for diagnosis of infectious diseases. Revue Scientifique et Technique (International Office of Epizootics) 17, 469526.
Jayaraj, R, Hales, B, Viberg, L, Pizzuto, S, Holt, D, Rolland, JM, O'Hehir, RE, Currie, BJ and Walton, SF (2011) A diagnostic test for scabies: IgE specificity for a recombinant allergen of Sarcoptes scabiei. Diagnostic Microbiology and Infectious Disease 71, 403407.
Ko, KM, Lee, W, Yu, JR and Ahnn, J (2007) PYP-1, inorganic pyrophosphatase, is required for larval development and intestinal function in C. elegans. FEBS Letters 581, 54455453.
Lacarrubba, F, Musumeci, ML, Caltabiano, R, Impallomeni, R, West, DP and Micali, G (2001) High-magnification videodermatoscopy: a new noninvasive diagnostic tool for scabies in children. Pediatric Dermatology 18, 439441.
Lahti, R, Kolakowski, LF, Heinonen, J, Vihinen, M, Pohjanoksa, K and Cooperman, BS (1990) Conservation of functional residues between yeast and E. coli inorganic pyrophosphatases. Biochimica et Biophysica Acta (BBA) – Protein Structure and Molecular Enzymology 1038, 338345.
Lemercier, G, Espiau, B, Ruiz, FA, Vieira, M, Luo, S, Baltz, T, Docampo, R and Bakalara, N (2004) A pyrophosphatase regulating polyphosphate metabolism in acidocalcisomes is essential for Trypanosoma brucei virulence in mice. Journal of Biological Chemistry 279, 34203425.
Ljunggren, E, Bergström, K, Morrison, D and Mattsson, J (2006) Characterization of an atypical antigen from Sarcoptes scabiei containing an MADF domain. Parasitology 132, 117126.
Lu, Y, Jia, R, Zhang, Z, Wang, M, Xu, Y, Zhu, D, Chen, S, Liu, M, Yin, Z and Chen, X (2014) In vitro expression and development of indirect ELISA for capsid protein of duck circovirus without nuclear localization signal. International Journal of Clinical and Experimental Pathology 7, 4938.
Mahmood, W, Viberg, LT, Fischer, K, Walton, SF and Holt, DC (2013) An aspartic protease of the scabies mite Sarcoptes scabiei is involved in the digestion of host skin and blood macromolecules. PLoS Neglected Tropical Diseases 7, e2525.
McDonald, M, Currie, BJ and Carapetis, JR (2004) Acute rheumatic fever: a chink in the chain that links the heart to the throat? The Lancet Infectious Diseases 4, 240245.
Micali, G, Lacarrubba, F and Lo, GG (1999) Scraping versus videodermatoscopy for the diagnosis of scabies: a comparative study. Acta Dermato-Venereologica 79, 396.
Micali, G, Lacarrubba, F and Tedeschi, A (2004) Videodermatoscopy enhances the ability to monitor efficacy of scabies treatment and allows optimal timing of drug application. Journal of the European Academy of Dermatology and Venereology 18, 153154.
Mimouni, D, Ankol, O, Davidovitch, N, Gdalevich, M, Zangvil, E and Grotto, I (2003) Seasonality trends of scabies in a young adult population: a 20-year follow-up. British Journal of Dermatology 149, 157159.
Morgan, MS and Arlian, LG (2006) Enzymatic activity in extracts of allergy-causing astigmatid mites. Journal of Medical Entomology 43, 12001207.
Ráez-Bravo, A, Granados, JE, Serrano, E, Dellamaria, D, Casais, R, Rossi, L, Puigdemont, A, Cano-Manuel, FJ, Fandos, P and Pérez, JM (2016) Evaluation of three enzyme-linked immunosorbent assays for sarcoptic mange diagnosis and assessment in the Iberian ibex, Capra pyrenaica. Parasites & Vectors 9, 558.
Rampton, M, Walton, SF, Holt, DC, Pasay, C, Kelly, A, Currie, BJ, McCarthy, JS and Mounsey, KE (2013) Antibody responses to Sarcoptes scabiei apolipoprotein in a porcine model: relevance to immunodiagnosis of recent infection. PLoS ONE 8, e65354.
Rider, SD Jr., Morgan, MS and Arlian, LG (2015) Draft genome of the scabies mite. Parasit Vectors 8, 585.
Rodríguez-Cadenas, F, Carbajal-González, M, Fregeneda-Grandes, J, Aller-Gancedo, J, Huntley, J and Rojo-Vázquez, F. A. (2010a). Development and evaluation of an antibody ELISA for sarcoptic mange in sheep and a comparison with the skin-scraping method. Preventive Veterinary Medicine 96, 8292.
Rodriguez-Cadenas, F, Carbajal-González, M, Fregeneda-Grandes, J, Aller-Gancedo, J. and Rojo-Vázquez, F. A. (2010b). Clinical evaluation and antibody responses in sheep after primary and secondary experimental challenges with the mange mite Sarcoptes scabiei var. ovis. Veterinary Immunology and Immunopathology 133, 109116.
Sanchez, J, Dohoo, IR, Markham, F, Leslie, K and Conboy, G (2002) Evaluation of the repeatability of a crude adult indirect Ostertagia ostertagi ELISA and methods of expressing test results. Veterinary Parasitology 109, 7590.
Sarasa, M, Rambozzi, L, Rossi, L, Meneguz, PG, Serrano, E, Granados, J.-E., González, FJ, Fandos, P, Soriguer, RC and Gonzalez, G (2010) Sarcoptes scabiei: specific immune response to sarcoptic mange in the Iberian ibex Capra pyrenaica depends on previous exposure and sex. Experimental Parasitology 124, 265271.
Steer, AC, Jenney, AW, Kado, J, Batzloff, MR, La Vincente, S, Waqatakirewa, L, Mulholland, EK and Carapetis, JR (2009) High burden of impetigo and scabies in a tropical country. PLoS Neglected Tropical Diseases 3, e467.
Tamura, K, Peterson, D, Peterson, N, Stecher, G, Nei, M and Kumar, S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28, 27312739.
Tarigan, S (2014) Antibody response in naïve and sensitised goats infested by Sarcoptes scabiei. Jurnal Ilmu Ternak dan Veteriner 9, 258265.
Thompson, JD, Gibson, TJ, Plewniak, F, Jeanmougin, F and Higgins, DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25, 48764882.
Van der Heijden, H, Rambags, P, Elbers, A, Van Maanen, C and Hunneman, W (2000) Validation of ELISAs for the detection of antibodies to Sarcoptes scabiei in pigs. Veterinary Parasitology 89, 95107.
Walton, SF and Currie, BJ (2007) Problems in diagnosing scabies, a global disease in human and animal populations. Clinical Microbiology Reviews 20, 268279.
Wilson, P, Slade, R, Currie, BJ, Walton, SF, Holt, DC, Fischer, K, Allen, GE, Wilson, D and Kemp, DJ (2003) Mechanisms for a novel immune evasion strategy in the scabies mite Sarcoptes scabiei: a multigene family of inactivated serine proteases. Journal of Investigative Dermatology 121, 14191424.
Worth, C, Heukelbach, J, Fengler, G, Walter, B, Liesenfeld, O and Feldmeier, H (2012) Impaired quality of life in adults and children with scabies from an impoverished community in Brazil. International Journal of Dermatology 51, 275282.
Xie, Y, Chen, S, Yan, Y, Zhang, Z, Li, D, Yu, H, Wang, C, Nong, X, Zhou, X and Gu, X (2013) Potential of recombinant inorganic pyrophosphatase antigen as a new vaccine candidate against Baylisascaris schroederi in mice. Veterinary Research 44, 90.
Yang, YY, Ko, TP, Chen, CC, Huang, G, Zheng, Y, Liu, W, Wang, I, Ho, MR, Hsu, S. T. D., O'Dowd, B, Huff, HC, Huang, CH, Docampo, R, Oldfield, E and Guo, RT (2016) Structures of trypanosome vacuolar soluble pyrophosphatases: antiparasitic drug targets. ACS Chemical Biology 11, 13621371.
Zhang, R, Zheng, W, Wu, X, Jise, Q, Ren, Y, Nong, X, Gu, X, Wang, S, Peng, X and Lai, S (2013) Characterisation and analysis of thioredoxin peroxidase as a potential antigen for the serodiagnosis of sarcoptic mange in rabbits by dot-ELISA. BMC Infectious Diseases 13, 336.
Zheng, Y, He, R, He, M, Gu, X, Wang, T, Lai, W, Peng, X and Yang, G (2016) Characterization of Sarcoptes scabiei cofilin gene and assessment of recombinant cofilin protein as an antigen in indirect-ELISA for diagnosis. BMC Infectious Diseases 16, 21.



Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed