SpringerLink
Log in

Evaluation of a PCR assay for diagnosis of toxoplasmosis in serum and peripheral blood mononuclear cell among HIV/AIDS patients

  • Original Article
  • Published:
Journal of Parasitic Diseases Aims and scope Submit manuscript

Abstract

Cerebral toxoplasmosis is one of the neurological infections with high morbidity and mortality in patients with AIDS, so the accurate method for diagnosis of cerebral toxoplasmosis seems necessary. In this study, nested PCR assay using B1 gene was evaluated in diagnosis of toxoplasmosis in serum and peripheral blood mononuclear cell (PBMC) among HIV/AIDS patients. One hundred eight blood samples from HIV/AIDS patients, including four patients with cerebral toxoplasmosis and 104 HIV/AIDS patients without cerebral toxoplasmosis were evaluated for the Toxoplasma gondii antibodies using Enzyme Linked immunosorbent Assay. DNA of serum and PBMC of these patients were extracted and nested-PCR was carried out. Of 108 participants, 95 cases (88%) were positive for Toxoplasma IgG antibodies and one patient was found positive for Toxoplasma IgM antibody. In general, four patients, including three patients with cerebral toxoplasmosis, who were positive for Toxoplasma IgG antibodies and one patient without cerebral toxoplasmosis who was positive for Toxoplasma IgM antibody were found to be PCR positive. DNA of T. gondii was detected in both serum and PBMC in two cerebral toxoplasmosis patients; however DNA was detected in only PBMC in other cerebral toxoplasmosis patient. All cases with cerebral toxoplasmosis were also diagnosed by clinical and radiological manifestations. The results of this study showed that the numbers of positive samples by PCR in PBMC were higher than serum specimens for diagnosis of toxoplasmosis. If molecular method and immunological assay are complemented with magnetic resonance imaging, the results can be useful for diagnosis of cerebral toxoplasmosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Ammassari A et al (2000) AIDS-related focal brain lesions in the era of highly active antiretroviral therapy. Neurology 55:1194–1200

    CAS  PubMed  Google Scholar 

  • Bourdin A et al (2014) PCR detection of Toxoplasma gondii DNA in blood and ocular samples for the diagnosis of ocular toxoplasmosis. J Clin Microbiol 52(11):3987–3991

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bretagne S, Costa JM, Foulet F, Jabot-Lestang L, Baud-Camus F, Cordonnier C (2000) Prospective study of toxoplasma reactivation by polymerase chain reaction in allogeneic stem-cell transplant recipients. Transpl Infect Dis 2:127–132

    CAS  PubMed  Google Scholar 

  • Burg JL, Grover CM, Pouletty P, Boothroyd J (1989) Direct and sensitive detection of a pathogenic protozoan, Toxoplasma gondii, by polymerase chain reaction. J Clin Microbiol 27:1787–1792

    CAS  PubMed  PubMed Central  Google Scholar 

  • Cardona N, Basto N, Parra B, Zea AF, Pardo CA, Bonelo A, Gómez-Marin JE (2011) Detection of Toxoplasma DNA in the peripheral blood of HIV-positive patients with neuro-opportunistic infections by a real-time PCR assay. J Neuroinfect Dis 2:1–6

    Google Scholar 

  • Cingolani A, De Luca A, Ammassari A, Murri R, Linzalone A, Grillo R, Antinori A (1996) PCR detection of Toxoplasma gondii DNA in CSF for the differential diagnosis of AIDS-related focal brain lesions. J Med Microbiol 45:472–476

    CAS  PubMed  Google Scholar 

  • Colombo FA et al (2005) Diagnosis of cerebral toxoplasmosis in AIDS patients in Brazil: importance of molecular and immunological methods using peripheral blood samples. J Clin Microbiol 43:5044–5047

    CAS  PubMed  PubMed Central  Google Scholar 

  • Contini C, Seraceni S, Cultrera R, Incorvaia C, Sebastiani A, Picot S (2005) Evaluation of a Real-time PCR-based assay using the lightcycler system for detection of Toxoplasma gondii bradyzoite genes in blood specimens from patients with toxoplasmic retinochoroiditis. Int J Parasitol 35:275–283

    CAS  PubMed  Google Scholar 

  • Derouin F et al (1996) Predictive value of Toxoplasma gondii antibody titres on the occurrence of toxoplasmic encephalitis in HIV-infected patients. ANRS 005/ACTG 154 Trial Group. AIDS 10:1521–1527

    CAS  PubMed  Google Scholar 

  • Donyavi T et al (2019) High prevalence of occult hepatitis C virus infection in injection drug users with HIV infection. Arch Virol 164:2493–2504

    CAS  PubMed  Google Scholar 

  • Dubey JP (2008) The history of Toxoplasma gondii—the first 100 years. J Eukaryot Microbiol 55:467–475

    PubMed  Google Scholar 

  • Dupouy-Camet J et al (1993) Detection of Toxoplasma gondii in venous blood from AIDS patients by polymerase chain reaction. J Clin Microbiol 31:1866–1869

    CAS  PubMed  PubMed Central  Google Scholar 

  • Esteghamati A, Khanaliha K, Bokharaei-Salim F, Sayyahfar S, Ghaderipour M (2019) Prevalence of Intestinal Parasitic Infection in Cancer, Organ Transplant and Primary Immunodeficiency Patients in Tehran, Iran. Asian Pac J Cancer Prev 20:495–501

    CAS  PubMed  PubMed Central  Google Scholar 

  • Fallahi S et al (2014) Comparison of the RE and B1 gene for detection of Toxoplasma gondii infection in children with cancer. Parasitol Int 63:37–41

    CAS  PubMed  Google Scholar 

  • Fricker-Hidalgo H et al (2009) Diagnosis of toxoplasmosis after allogeneic stem cell transplantation: results of DNA detection and serological techniques. Clin Infect Dis 48:e9–e15

    PubMed  Google Scholar 

  • Gashout A et al (2016) Molecular diagnosis of Toxoplasma gondii infection in Libya. BMC Infect Dis 16:157

    PubMed  PubMed Central  Google Scholar 

  • Gianotti N, Cinque P, Castagna A, Novati R, Moro M, Lazzarin A (1997) Diagnosis of toxoplasmic encephalitis in HIV-infected patients. AIDS 11:1529

    CAS  PubMed  Google Scholar 

  • Hellerbrand C, Goebel F, Disko R (1996) High predictive value of Toxoplasma gondii IgG antibody levels in HIV-infected patients for diagnosis of cerebral toxoplasmosis. Eur J Clin Microbiol Infect Dis 15:869–872

    CAS  PubMed  Google Scholar 

  • Holliman RE (1988) Toxoplasmosis and the acquired immune deficiency syndrome. J Infect 16:121–128

    CAS  PubMed  Google Scholar 

  • Jones CD, Okhravi N, Adamson P, Tasker S, Lightman S (2000) Comparison of PCR detection methods for B1, P30, and 18S rDNA genes of T. gondii in aqueous humor. Invest Ophthalmol Vis Sci 41:634–644

    CAS  PubMed  Google Scholar 

  • Joseph P et al (2002) Optimization and evaluation of a PCR assay for detecting toxoplasmic encephalitis in patients with AIDS. J Clin Microbiol 40:4499–4503

    CAS  PubMed  PubMed Central  Google Scholar 

  • Khanaliha K, Motazedian M, Sarkari B, Bandehpour M, Sharifnia Z, Kazemi B (2012) Expression and purification of P43 Toxoplasma gondii surface antigen. Iran J Parasitol 7:48

    CAS  PubMed  PubMed Central  Google Scholar 

  • Khanaliha K, Motazedian MH, Kazemi B, Shahriari B, Bandehpour M, Sharifniya Z (2014) Evaluation of recombinant SAG1, SAG2, and SAG3 antigens for serodiagnosis of toxoplasmosis. Korean J Parasitol 52:137

    CAS  PubMed  PubMed Central  Google Scholar 

  • Khanaliha K, Mohebali M, Davoudi S, Hosseini OR, Tarighi F, Rezaeian T, Rezaeian M (2015) Detection of emergence Cyclospora cayetanensis in A HIV+/AIDS patient with diarrhea from Tehran: a case report. Iran J Public Health 44:865

    PubMed  PubMed Central  Google Scholar 

  • Lamoril J, Molina J, De Gouvello A, Garin Y, Deybach J, Modai J, Derouin F (1996) Detection by PCR of Toxoplasma gondii in blood in the diagnosis of cerebral toxoplasmosis in patients with AIDS. J Clin Pathol 49:89–92

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lee S-B, Lee T-G (2017) Toxoplasmic encephalitis in patient with acquired immunodeficiency syndrome. Brain Tumor Res Treat 5:34–36

    PubMed  PubMed Central  Google Scholar 

  • Luft BJ, Chua A (2000) Central nervous system toxoplasmosis in HIV pathogenesis, diagnosis, and therapy. Curr Infect Dis Rep 2:358–362

    CAS  PubMed  Google Scholar 

  • Luft BJ, Brooks RG, Conley FK, McCabe RE, Remington JS (1984) Toxoplasmic encephalitis in patients with acquired immune deficiency syndrome. JAMA 252:913–917

    CAS  PubMed  Google Scholar 

  • Masoumi-Asl H, Khanaliha K, Bokharaei-Salim F, Esteghamati A, Kalantari S, Hosseinyrad M (2019) Enteric Opportunistic Infection and the Impact of Antiretroviral Therapy among HIV/AIDS Patients from Tehran, Iran. Iran J Public Health 48:730

    PubMed  PubMed Central  Google Scholar 

  • Nissapatorn V (2008) Lessons learned about opportunistic infections in southeast Asia. Southeast Asian J Trop Med Public Health 39:625

    PubMed  Google Scholar 

  • Novati R et al (1994) Polymerase chain reaction for Toxoplasma gondii DNA in the cerebrospinal fluid of AIDS patients with focal brain lesions. AIDS 8:1691–1694

    CAS  PubMed  Google Scholar 

  • Pelloux H, Dupouy-Camet J, Derouin F, Aboulker J, Raffi F, Group B-TS (1997) A multicentre prospective study for the polymerase chain reaction detection of Toxoplasma gondii DNA in blood samples from 186 AIDS patients with suspected toxoplasmic encephalitis. AIDS 11:1888–1890

    CAS  PubMed  Google Scholar 

  • Rostami A, Keshavarz H, Shojaee S, Mohebali M, Meamar AR (2014) Frequency of Toxoplasma gondii in HIV positive patients from West of Iran by ELISA and PCR. Iran J Parasitol 9:474

    PubMed  PubMed Central  Google Scholar 

  • Rostami A, Karanis P, Fallahi S (2018) Advances in serological, imaging techniques and molecular diagnosis of Toxoplasma gondii infection. Infection 46:303–315

    CAS  PubMed  Google Scholar 

  • Sacktor N (2002) The epidemiology of human immunodeficiency virus-associated neurological disease in the era of highly active antiretroviral therapy. J Neurovirol 8(Suppl 2):115–121

    CAS  PubMed  Google Scholar 

  • Sadler M, Brink N, Gazzard B (1998) Management of intracerebral lesions in patients with HIV: a retrospective study with discussion of diagnostic problems. QJM 91:205–217

    CAS  PubMed  Google Scholar 

  • Sayyahfar S, Karimi A, Gharib A, Fahimzad A (2015) Association of systemic anaplastic large cell lymphoma and active toxoplasmosis in a child. Iran J Cancer Prev 8(4):e3438

    PubMed  PubMed Central  Google Scholar 

  • Sensini A (2006) Toxoplasma gondii infection in pregnancy: opportunities and pitfalls of serological diagnosis. Clin Microbiol Infect 12:504–512

    CAS  PubMed  Google Scholar 

  • Shiadeh MN et al (2016) The correlation between Toxoplasma gondii infection and prenatal depression in pregnant women. Eur J Clin Microbiol Infect Dis 35:1829–1835

    Google Scholar 

  • Silveira C et al (2011) Toxoplasma gondii in the peripheral blood of patients with acute and chronic toxoplasmosis. Br J Ophthalmol 95:396–400

    PubMed  Google Scholar 

  • Vidal JE, Colombo FA, de Oliveira ACP, Focaccia R, Pereira-Chioccola VL (2004) PCR assay using cerebrospinal fluid for diagnosis of cerebral toxoplasmosis in Brazilian AIDS patients. J Clin Microbiol 42:4765–4768

    PubMed  PubMed Central  Google Scholar 

  • Wahab T, Edvinsson B, Palm D, Lindh J (2010) Comparison of the AF146527 and B1 repeated elements, two real-time PCR targets used for detection of Toxoplasma gondii. J Clin Microbiol 48:591–592

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors would like to thank from staffs of Infectious Diseases ward, Rasoul-e-Akram Hospital, Iran University of Medical Sciences for their helpful cooperation.

Funding

This study was funded by Research Center of Pediatric Infectious Diseases, Iran University of Medical Sciences in Tehran, Iran with Grant Number (95-02-131-28979)

Author information

Authors and Affiliations

  1. Department of Virology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Farah Bokharaei-Salim

  2. Research Center of Pediatric Infectious Diseases, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences, Tehran, Iran

    Abdoulreza Esteghamati, Khadijeh Khanaliha & Shirin Sayyahfar

  3. Department of Infectious Diseases, Rasoul-e-Akram Hospital, Iran University of Medical Sciences, Tehran, Iran

    Saeed Kalantari

  4. Iran University of Medical Sciences, Tehran, Iran

    Tahereh Donyavi & Saba Garshasbi

  5. Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

    Qasem Asgari

  6. Student Research Committee, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

    Borna Salemi

Authors
  1. Farah Bokharaei-Salim
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Abdoulreza Esteghamati
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Khadijeh Khanaliha
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Saeed Kalantari
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shirin Sayyahfar
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tahereh Donyavi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Saba Garshasbi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Qasem Asgari
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Borna Salemi
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

KK and FBS proposed the study. KK, AE and SK designed the study. TD, SG and QA help in technical works and Borna Salami helps in sample collection. KK and FBS and SS revised the manuscript. All the authors approved the final manuscript

Corresponding author

Correspondence to Khadijeh Khanaliha.

Ethics declarations

Ethic approval

Ethical approval for the study and informed consent forms were approved by ethics committee of Iran University of Medical Sciences (Code Number: IR.IUMS.REC 1395.95-02-131-28979). Ethical approval for this study was obtained from IUMS’s Ethics Committee, and written informed consent was taken from all patients or Legally Authorized Representative. All patients or relatives informed the study procedures and agreed to participate in this study.

Conflict of interest

The authors declare that they have no conflicts of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bokharaei-Salim, F., Esteghamati, A., Khanaliha, K. et al. Evaluation of a PCR assay for diagnosis of toxoplasmosis in serum and peripheral blood mononuclear cell among HIV/AIDS patients. J Parasit Dis 44, 159–165 (2020). https://doi.org/10.1007/s12639-019-01176-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12639-019-01176-6

Keywords

Search

Navigation