Strongyloides stercoralis: detection of parasite-derived DNA in serum samples obtained from immunosuppressed patients
Tóm tắt
Strongyloidiasis is an important neglected disease, which is life threatening in immunocompromised patients. This study aimed to evaluate the frequency of Strongyloides stercoralis infection among immunosuppressed subjects living in endemic communities by conventional PCR of the 18S rRNA and Cox1 genes to detect cell-free DNA in the patients’ serum samples. Fresh stool and serum samples were obtained from participants. The stool samples were examined using parasitological methods. Total DNA was extracted from the serum samples and S. stercoralis larvae isolated from patient fecal samples. Conventional PCR to amplify a 101 bp fragment of the 18S rRNA gene was carried out for all extracted DNA, and then positive samples were further evaluated for a 121 bp fragment of the Cox1 gene. The PCR products of selected samples were sequenced and BLAST analysis was performed. Out of 120 patients, 57 and 63 cases had autoimmune disorders and cancer, respectively. The 101 bp fragments of the 18S rRNA were successfully amplified in 36 out of 120 (30%) serum samples. The PCR products of five samples were sequenced and compared with reference sequences in GenBank, which showed 97% identity and 90% coverage. In conclusion, to the best of our knowledge, this is the first molecular study for the detection of S. stercoralis cell-free DNA in human serum samples. These results provide useful insights for future studies and show that serum is an alternative specimen and may be useful in molecular diagnosis of diseases, particularly in immunosuppressive patients.
Tài liệu tham khảo
Beknazarova M, Whiley H, Ross K (2016) Strongyloidiasis: a disease of socioeconomic disadvantage. Int J Environ Res Public Health 13. https://doi.org/10.3390/ijerph13050517
Buonfrate D, Formenti F, Perandin F, Bisoffi Z (2015) Novel approaches to the diagnosis of Strongyloides stercoralis infection. Clin Microbiol Infect 21:543–552. https://doi.org/10.1016/j.cmi.2015.04.001
Dorris M, Viney ME, Blaxter ML (2002) Molecular phylogenetic analysis of the genus Strongyloides and related nematodes. Int J Parasitol 32:1507–1517. https://doi.org/10.1016/S0020-7519(02)00156-X
Ek T, Mellander L, Hahn-Zoric M, Abrahamsson J (2006) Avidity of tetanus and Hib antibodies after childhood acute lymphoblastic leukaemia—implications for vaccination strategies. Acta Paediatr 95:701–706
Fernández-Soto P et al (2016) Strong-LAMP: a LAMP assay for Strongyloides spp. detection in stool and urine samples. Towards the diagnosis of human strongyloidiasis starting from a rodent model. PLoS Negl Trop Dis 10:e0004836. https://doi.org/10.1371/journal.pntd.0004836
Hasegawa H, Sato H, Fujita S, Nguema PPM, Nobusue K, Miyagi K, Kooriyama T, Takenoshita Y, Noda S, Sato A, Morimoto A, Ikeda Y, Nishida T (2010) Molecular identification of the causative agent of human strongyloidiasis acquired in Tanzania: dispersal and diversity of Strongyloides spp. and their hosts. Parasitol Int 59:407–413. https://doi.org/10.1016/j.parint.2010.05.007
Hasegawa H, Kalousova B, McLennan MR, Modry D, Profousova-Psenkova I, Shutt-Phillips KA, Todd A, Huffman MA, Petrzelkova KJ (2016) Strongyloides infections of humans and great apes in Dzanga-Sangha Protected Areas, Central African Republic and in degraded forest fragments in Bulindi, Uganda. Parasitol Int 65:367–370. https://doi.org/10.1016/j.parint.2016.05.004
Jaleta TG, Zhou S, Bemm FM, Schär F, Khieu V, Muth S, Odermatt P, Lok JB, Streit A (2017) Different but overlapping populations of Strongyloides stercoralis in dogs and humans—dogs as a possible source for zoonotic strongyloidiasis. PLOS Neg Trop Dis 11:e0005752. https://doi.org/10.1371/journal.pntd.0005752
Janwan P, Intapan PM, Thanchomnang T, Lulitanond V, Anamnart W, Maleewong W (2011) Rapid detection of Opisthorchis viverrini and Strongyloides stercoralis in human fecal samples using a duplex real-time PCR and melting curve analysis. Parasitol Res 109:1593–1601. https://doi.org/10.1007/s00436-011-2419-z
Keiser PB, Nutman TB (2004) Strongyloides stercoralis in the immunocompromised population. Clin Microbiol Rev 17:208–217. https://doi.org/10.1128/cmr.17.1.208-217.2004
Kia EB, Mahmoudi M, Zahabiun F, Meamar AR (2007) An evaluation on the efficacy of agar plate culture for detection of Strongyloides stercoralis. Iran J Parasitol 2:29–34
Kramme S, Nissen N, Soblik H, Erttmann K, Tannich E, Fleischer B, Panning M, Brattig N (2011) Novel real-time PCR for the universal detection of Strongyloides species. J Med Microbiol 60:454–458. https://doi.org/10.1099/jmm.0.025338-0
Lodh N, Caro R, Sofer S, Scott A, Krolewiecki A, Shiff C (2016) Diagnosis of Strongyloides stercoralis: detection of parasite-derived DNA in urine. Acta Trop 163:9–13. https://doi.org/10.1016/j.actatropica.2016.07.014
Machado ER, Teixeira EM, Gonçalves-pires MF, Loureiro ZM, Araújo RA, Costa-cruz JM (2008) Parasitological and immunological diagnosis of Strongyloides stercoralis in patients with gastrointestinal cancer. Scand J Infect Dis 40:154–158. https://doi.org/10.1080/00365540701558730
Mejia R, Nutman TB (2012) Screening, prevention, and treatment for hyperinfection syndrome and disseminated infections caused by Strongyloides stercoralis. Curr Opin Infect Dis 25:458–463. https://doi.org/10.1097/QCO.0b013e3283551dbd
Mendes T, Minori K, Ueta M, Miguel DC, Allegretti SM (2017) Strongyloidiasis current status with emphasis in diagnosis and drug research. J Parasitol Res 2017. https://doi.org/10.1155/2017/5056314
Moghaddassani H, Mirhendi H, Hosseini M, Rokni MB, Mowlavi G, Kia E (2011) Molecular diagnosis of Strongyloides stercoralis infection by PCR detection of specific DNA in human stool samples. Iran J Parasitol 6:23–30
Nutman TB (2017) Human infection with Strongyloides stercoralis and other related Strongyloides species. Parasitology 144:263–273. https://doi.org/10.1017/s0031182016000834
Paula FM, Malta FM, Corral MA, Marques PD, Gottardi M, Meisel DM Yamashiro J, Pinho JR, Castilho VL, Goncalves EM, Gryschek RC, Chieffi PP (2016) Diagnosis of Strongyloides stercoralis infection in immunocompromised patients by serological and molecular methods. Rev Inst Med Trop Sao Paulo 58:c. https://doi.org/10.1590/s1678-9946201658063
Putland RA, Thomas SM, Grove DI, Johnson AM (1993) Analysis of the 18S ribosomal RNA gene of Strongyloides stercoralis. Int J Parasitol 23:149–151
Salvador F, Sulleiro E, Sánchez-Montalvá A, Saugar JM, Rodríguez E, Pahissa A, Molina I (2014) Usefulness of Strongyloides stercoralis serology in the management of patients with eosinophilia. Am J Trop Med Hyg 90:830–834. https://doi.org/10.4269/ajtmh.13-0678
Sato Y, Kobayashi J, Shiroma Y (1995a) Serodiagnosis of strongyloidiasis. The application and significance. Rev Inst Med Trop Sao Paulo 37:35–41
Sato Y, Kobayashi J, Toma H, Shiroma Y (1995b) Efficacy of stool examination for detection of Strongyloides infection. Am J Trop Med Hyg 53:248–250
Siddiqui AA, Berk SL (2001) Diagnosis of Strongyloides stercoralis infection. Clin Infect Dis 33:1040–1047. https://doi.org/10.1086/322707
Verweij JJ, Canales M, Polman K, Ziem J, Brienen EAT, Polderman AM, van Lieshout L (2009) Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR. Trans R Soc Trop Med Hyg 103:342–346. https://doi.org/10.1016/j.trstmh.2008.12.001
Verweij JJ, Canales M, Polman K, Ziem J, Brienen EAT, Polderman AM, van Lieshout L (2010) Corrigendum to “Molecular diagnosis of Strongyloides stercoralis in faecal samples using real-time PCR” [Transactions of the Royal Society of Tropical Medicine and Hygiene (2009) 103, 342–6]. Trans R Soc Trop Med Hyg 104:378. https://doi.org/10.1016/j.trstmh.2010.02.010
Zueter AM, Mohamed Z, Abdullah AD, Mohamad N, Arifin N, Othman N, Noordin R (2014) Detection of Strongyloides stercoralis infection among cancer patients in a major hospital in Kelantan, Malaysia. Singap Med J 55:367–371