Molecular and Immnune Diagnosis: Further Testing for Human Strongyloidiasis
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Detection of Strongyloides stercoralis larvae is particularly challenging because only a small number of larvae are released into the feces, regardless of infection stage.
Our objective was to apply conventional polymerase chain reaction (PCR) to the detection of S. stercoralis DNA in feces samples to evaluate its performance in samples of patients with strongyloidiasis and compare results with those of immunodiagnosis.
Stool, serum, and saliva samples were collected from each individual (n = 48) at the clinic hospital of the State University of Londrina, Brazil, for parasitological, immunological, and molecular tests. Stool samples were processed via parasitological methods. Serum samples were used for immunoglobulin G (IgG) detection and saliva samples for IgA detection by ELISA.
For amplification by conventional PCR, two different primers were used: species specific (101 bp) and genus specific (392 bp). The results showed that 34 (97.1%) of the 35 copro-positive individuals for S. stercoralis were positive for serum IgG and 19 (54.3%) were positive for salivary IgA. Regarding molecular analysis, both primers (species and genus specific) demonstrated positivity in 100% of the samples, which was confirmed by sequencing the positive samples.
Complementary examinations of the parasitological method demonstrated excellent results in the context of the diagnosis of strongyloidiasis, especially in asymptomatic patients with irregular larval release in the feces.
Compliance with Ethical Standards
This study was supported by grants from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES—Grant no. 40002012026M9 [LRB] and Fundação de Amparo a Pesquisa do estado de São Paulo (FAPESP—Grant no. 2013/03304-0).
Conflict of interest
LRB, PDM, GBdM, MdRFG-P, FMM, WRP, IC-C, JMC-C, FMP and INC have no conflicts of interest.
Ethical Approval and Informed Consent
The study was approved by the ethics committee on human studies at the State University of Londrina (protocol number 1.306.715).
- 1.Buonfrate D, Sequi M, Mejia R, Cimino RO, Krolewiecki AJ, Albonico M, Degani M, Tais S, Angheben A, Requena-Mendez A, Muñoz J, Nutman TB, Bisoffi Z. Accuracy of five serologic tests for the follow up of Strongyloides stercoralis infection. PLoS Negl Trop Dis. 2015;9:e0003491.CrossRefPubMedPubMedCentralGoogle Scholar
- 6.Yap P, Fürst T, Müller I, Kriemler S, Utzinger J, Steinmann P. Determining soil-transmitted helminth infection status and physical fitness of school-aged children. J Vis Exp. 2012;66:e3966.Google Scholar
- 7.Llewellyn S, Inpankaew T, Nery SV, Gray DJ, Verweij JJ, Clements AC, Gomes SJ, Traub R, McCarthy JS. Application of a multiplex quantitative PCR to assess prevalence and intensity of intestinal parasite infections in a controlled clinical trial. PLoS Negl Trop Dis. 2016;10:e0004380.CrossRefPubMedPubMedCentralGoogle Scholar
- 10.Nilforoushan MR, Mirhendi H, Rezaie S, Rezaian M, Meamar AR, Kia EB. A DNA-based identification of Strongyloides stercoralis isolates from Iran. Iran J Publ Health. 2007;36:16–20.Google Scholar
- 14.Paula FM, Malta FM, Marques PD, Sitta RB, Pinho JR, Gryschek RCB, Chieffi PP. Molecular diagnosis of strongyloidiasis in tropical areas: a comparison of conventional and real-time polymerase chain reaction with parasitological methods. Mem. Inst. Oswaldo Cruz 2015;110:272–4.Google Scholar
- 17.Paula FM, Malta FM, Corral MA, Marques PD, Gottardi M, Meisel DM, Yamashiro J, Pinho JR, Castilho VL, Gonçalves EM, Gryschek RC, Chieffi PP. Diagnosis of Strongyloides stercoralis infection in immunocompromised patients by serological and molecular methods. Rev Inst Med Trop Sao Paulo 2016;58:63.Google Scholar
- 18.Hoffmann WA, Pons JA, Janer JL. The sedimentation concentration method in schistosomiasis mansoni, Puerto Rico. J Publ Health Trop Med. 1934;9:283–91.Google Scholar
- 20.Kato K, Miura M. Comparative examinations. Jpn J Parasitol. 1954;3:35.Google Scholar
- 23.Costa IN, Sopelete MC, Gonçalves-Pires MRF, Costa-Cruz JM. IgA and IgG antibodies in paired serum and saliva samples in human strongyloidiasis. Acta Parasitol. 2003;48:306–11.Google Scholar
- 28.Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci EUA 1977; 74:5463–7.Google Scholar
- 34.Bosqui LR, Gonçalves AL, Gonçalves-Pires Mdo R, Custodio LA, de Menezes MC, Murad VA, de Paula FM, Pavanelli WR, Conchon-Costa I, Costa-Cruz JM, Costa IN. Detection of parasite-specific IgG and IgA in paired serum and saliva samples for diagnosis of human strongyloidiasis in northern Paraná state, Brazil. Acta Trop. 2015;150:190–5.CrossRefPubMedGoogle Scholar
- 41.Khare R, Espy MJ, Cebelinski E, Boxrud D, Sloan LM, Cunningham SA, Pritt BS, Patel R, Binnicker MJ. Comparative evaluation of two commercial multiplex panels for detection of gastrointestinal pathogens by use of clinical stool specimens. J Clin Microbiol. 2014;52:3667–73.CrossRefPubMedPubMedCentralGoogle Scholar
- 42.Buonfrate D, Requena-Mendez A, Angheben A, Cinquini M, Cruciani M, Fittipaldo A, Giorli G, Gobbi F, Piubelli C, Bisoffi Z. Accuracy of molecular biology techniques for the diagnosis of Strongyloides stercoralis infection—a systematic review and meta-analysis. PLoS Negl Trop Dis. 2018;12:e0006229.CrossRefPubMedPubMedCentralGoogle Scholar