Evaluation of rK-39 Strip Test Using Urine for Diagnosis of Visceral Leishmaniasis in an Endemic Region of India

Dharmendra Singh Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Dharmendra Singh in
Current site
Google Scholar
PubMed
Close
,
Krishna Pandey Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Krishna Pandey in
Current site
Google Scholar
PubMed
Close
,
Vidya Nand Rabi Das Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Vidya Nand Rabi Das in
Current site
Google Scholar
PubMed
Close
,
Sushmita Das Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Sushmita Das in
Current site
Google Scholar
PubMed
Close
,
Neena Verma Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Neena Verma in
Current site
Google Scholar
PubMed
Close
,
Alok Ranjan Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Alok Ranjan in
Current site
Google Scholar
PubMed
Close
,
Sekhar Chandra Lal Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Sekhar Chandra Lal in
Current site
Google Scholar
PubMed
Close
,
Kamal Roshan Topno Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Kamal Roshan Topno in
Current site
Google Scholar
PubMed
Close
,
Shubhankar Kumar Singh Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Shubhankar Kumar Singh in
Current site
Google Scholar
PubMed
Close
,
Rakesh Bihari Verma Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Rakesh Bihari Verma in
Current site
Google Scholar
PubMed
Close
,
Ashish Kumar Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Ashish Kumar in
Current site
Google Scholar
PubMed
Close
,
Abul Hasan Sardar Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Abul Hasan Sardar in
Current site
Google Scholar
PubMed
Close
,
Bidyut Purkait Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Bidyut Purkait in
Current site
Google Scholar
PubMed
Close
, and
Pradeep Das Departments of Molecular Biology, Clinical Medicine, Pathology, Biostatistics, Biochemistry, Epidemiology, and Immunology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India

Search for other papers by Pradeep Das in
Current site
Google Scholar
PubMed
Close
Restricted access

The definitive diagnosis of visceral leishmaniasis (VL) requires invasive procedures for demonstration of parasites in tissue smear or culture. These procedures need expertise and laboratory supports and cannot be performed in the field. The aim of the present study was to evaluate the existing rK-39 immunochromatographic nitrocellulose strips test (ICT) with some modification in human urine for diagnosis of VL. The test was performed on both sera and urine samples on the same 786 subjects (365 confirmed VL and 421 control subjects). The sensitivity of the rK-39 ICT in serum was 100%, whereas the specificity was 93.8%, 100%, and 96.2% in healthy controls from endemic, non-endemic, and other infectious diseases, respectively. However, in urine samples, the test showed 96.1% sensitivity and 100% specificity. Considering sensitivity and feasibility of the test in the field, rK-39 ICT using urine samples can be an alternative to conventional invasive VL diagnosis.

Author Notes

* Address correspondence to Pradeep Das, Department of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna—800 007, Bihar, India. E-mail: drpradeep.das@gmail.com
† These authors contributed equally.

Authors' addresses: Dharmendra Singh, Sushmita Das, Ashish Kumar, Abul Hasan Sardar, Bidyut Purkait, and Pradeep Das, Department of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mails: singhd72@yahoo.co.in, sushmita.de2008@gmail.com, ashish2k8@gmail.com, abulhasansardar82@gmail.com, bidybiotech@gmail.com, and rmrims@icmr.org.in. Krishna Pandey, Vidya Nand Rabi Das, and Rakesh Bihari Verma, Department of Clinical Medicine, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mails: drkrishnapandey@yahoo.com, drvnrdas@yahoo.com, and rbihariverma@yahoo.com. Neena Verma, Department of Pathology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mail: verma_neena@yahoo.com. Alok Ranjan, Department of Health Research Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mail: aranjan30@yahoo.com. Sekhar Chandra Lal, Department of Biochemistry, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mail: drcslal@sify.com. Kamal Roshan Topno, Department of Epidemiology, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mail: roshanktopno@yahoo.co.in. Shubhankar Kumar Singh, Immunology Division, Rajendra Memorial Research Institute of Medical Sciences (Indian Council of Medical Research), Agamkuan, Patna, Bihar, India, E-mail: sksrmri@yahoo.com.

  • 1.

    Lukes J, Mauricio IL, Schönian G, Dujardin JC, Soteriadou K, Dedet JP, Kuhls K, Tintaya KW, Jirků M, Chocholová E, Haralambous C, Pratlong F, Oborník M, Horák A, Ayala FJ, Miles MA, 2007. Evolutionary and geographical history of the Leishmania donovani complex with a revision of current taxonomy. Proc Natl Acad Sci USA 104: 93759380.

    • Search Google Scholar
    • Export Citation
  • 2.

    Berman JD, 1997. Human leishmaniasis: clinical, diagnostic, and chemotherapeutic developments in the last 10 years. Clin Infect Dis 24: 684703.

    • Search Google Scholar
    • Export Citation
  • 3.

    Desjeux P, 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27: 305318.

  • 4.

    Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, Jannin J, den Boer M; the WHO Leishmaniasis Control Team, 2012. Leishmaniasis worldwide and global estimates of its incidence. PLoS One 7: e35671.

    • Search Google Scholar
    • Export Citation
  • 5.

    Stauch A, Sarkar RR, Picado A, Ostyn B, Sundar S, Rijal S, Boelaert M, Dujardin JC, Duerr HP, 2011. Visceral leishmaniasis in the Indian subcontinent: modelling epidemiology and control. PLoS Negl Trop Dis 5: e1405.

    • Search Google Scholar
    • Export Citation
  • 6.

    Das P, Samuels S, Desjeux P, Mittal A, Topno R, Siddiqui NA, Sur D, Pandey A, Sarnoff R, 2010. Annual incidence of visceral leishmaniasis in an endemic area of Bihar, India. Trop Med Int Health 15 (Suppl 2): 411.

    • Search Google Scholar
    • Export Citation
  • 7.

    Srividya G, Kulshrestha A, Singh R, Salotra P, 2012. Diagnosis of visceral leishmaniasis: developments over the last decade. Parasitol Res 110: 10651078.

    • Search Google Scholar
    • Export Citation
  • 8.

    Zijlstra EE, Ali MS, el-Hassan AM, el-Toum IA, Satti M, Ghalib HW, Kager PA, 1992. Kala-azar: a comparative study of parasitological methods and the direct agglutination test in diagnosis. Trans R Soc Trop Med Hyg 86: 505507.

    • Search Google Scholar
    • Export Citation
  • 9.

    Desjeux P, 1996. Leishmaniasis. Public health aspects and control. Clin Dermatol 14: 417423.

  • 10.

    Kumar S, Kumar D, Chakravarty J, Rai M, Sundar S, 2012. Identification and characterization of a novel Leishmania donovani antigen for serodiagnosis of visceral leishmaniasis. Am J Trop Med Hyg 86: 601605.

    • Search Google Scholar
    • Export Citation
  • 11.

    Pedras MJ, de Gouvêa Viana L, de Oliveira EJ, Rabello A, 2008. Comparative evaluation of direct agglutination test, rK39 and soluble antigen ELISA and IFAT for the diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg 102: 172178.

    • Search Google Scholar
    • Export Citation
  • 12.

    Gidwani K, Kumar R, Rai M, Sundar S, 2009. Longitudinal seroepidemiologic study of visceral leishmaniasis in hyperendemic regions of Bihar, India. Am J Trop Med Hyg 80: 345346.

    • Search Google Scholar
    • Export Citation
  • 13.

    Fisa R, Riera C, López-Chejade P, Molina I, Gállego M, Falcó V, Ribera E, Portús M, 2008. Leishmania infantum DNA detection in urine from patients with visceral leishmaniasis and after treatment control. Am J Trop Med Hyg 78: 741744.

    • Search Google Scholar
    • Export Citation
  • 14.

    Mary C, Faraut F, Lascombe L, Dumon H, 2004. Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. J Clin Microbiol 42: 52495255.

    • Search Google Scholar
    • Export Citation
  • 15.

    Salotra P, Sreenivas G, Pogue GP, Lee N, Nakhasi HL, Ramesh V, Negi NS, 2001. Development of a species-specific PCR assay for detection of Leishmania donovani in clinical samples from patients with kala-azar and post-kala-azar dermal leishmaniasis. J Clin Microbiol 39: 849854.

    • Search Google Scholar
    • Export Citation
  • 16.

    Srivastava P, Mehrotra S, Tiwary P, Chakravarty J, Sundar S, 2011. Diagnosis of Indian visceral leishmaniasis by nucleic acid detection using PCR. PLoS One 6: e19304.

    • Search Google Scholar
    • Export Citation
  • 17.

    de Assis TS, Braga AS, Pedras MJ, Oliveira E, Barral A, de Siqueira IC, Costa CH, Costa DL, Holanda TA, Soares VY, Biá M, Caldas A de J, Romero GA, Rabello A, 2010. Multi-centric prospective evaluation of rk39 rapid test and direct agglutination test for the diagnosis of visceral leishmaniasis in Brazil. Trans R Soc Trop Med Hyg 105: 8185.

    • Search Google Scholar
    • Export Citation
  • 18.

    Maia Z, Lírio M, Mistro S, Mendes CM, Mehta SR, Badaro R, 2012. Comparative study of rK39 Leishmania antigen for serodiagnosis of visceral leishmaniasis: systematic review with meta-analysis. PLoS Negl Trop Dis 6: e1484.

    • Search Google Scholar
    • Export Citation
  • 19.

    Sundar S, Reed SG, Singh VP, Kumar PC, Murray HW, 1998. Rapid accurate field diagnosis of Indian visceral leishmaniasis. Lancet 351: 563565.

  • 20.

    Zijlstra EE, Nur Y, Desjeux P, Khalil EA, El Hassan AM, Groen J, 2001. Diagnosing visceral leishmaniasis with the recombinant K39 strip test: experience from the Sudan. Trop Med Int Health 6: 108113.

    • Search Google Scholar
    • Export Citation
  • 21.

    Mondal D, Singh SP, Kumar N, Joshi A, Sundar S, Das P, Siddhivinayak H, Kroeger A, Boelaert M, 2009. Visceral leishmaniasis elimination program in India, Bangladesh, and Nepal: reshaping the case finding/case management strategy. PLoS Negl Trop Dis 3: e355.

    • Search Google Scholar
    • Export Citation
  • 22.

    WHO/SEARO, 2005. Regional Strategic Framework for Elimination of Kala-Azar from the South-East Asia Region (2005–2015). Geneva: WHO/SEARO.

    • Search Google Scholar
    • Export Citation
  • 23.

    Bhattacharya SK, Sur D, Sinha PK, Karbwang J, 2006. Elimination of leishmaniasis (kala-azar) from the Indian subcontinent is technically feasible and operationally achievable. Indian J Med Res 123: 195196.

    • Search Google Scholar
    • Export Citation
  • 24.

    Hirve S, Singh SP, Kumar N, Banjara MR, Das P, Sundar S, Rijal S, Joshi A, Kroeger A, Varghese B, Thakur CP, Huda MM, Mondal D, 2010. Effectiveness and feasibility of active and passive case detection in the visceral leishmaniasis elimination initiative in India, Bangladesh, and Nepal. Am J Trop Med Hyg 83: 507511.

    • Search Google Scholar
    • Export Citation
  • 25.

    Thakur CP, Meenakshi Thakur AK, Thakur S, 2009. Newer strategies for the kala-azar elimination programme in India. Indian J Med Res 129: 102104.

    • Search Google Scholar
    • Export Citation
  • 26.

    Matlashewski G, Arana B, Kroeger A, Battacharya S, Sundar S, Das P, Sinha PK, Rijal S, Mondal D, Zilberstein D, Alvar J, 2011. Visceral leishmaniasis: elimination with existing interventions. Lancet Infect Dis 11: 322325.

    • Search Google Scholar
    • Export Citation
  • 27.

    Saha S, Goswami R, Pramanik N, Guha SK, Saha B, Rahman M, Mallick S, Modak D, Silva FO, Mendonca IL, Costa DL, Costa CH, Ali N, 2011. Easy test for visceral leishmaniasis and post-kala-azar dermal leishmaniasis. Emerg Infect Dis 17: 13041306.

    • Search Google Scholar
    • Export Citation
  • 28.

    Galaï Y, Chabchoub N, Ben-Abid M, Ben-Abda I, Ben-Alaya-Bouafif N, Amri F, Aoun K, Bouratbine A, 2011. Diagnosis of Mediterranean visceral leishmaniasis by detection of Leishmania antibodies and Leishmania DNA in oral fluid samples collected using an Oracol device. J Clin Microbiol 49: 31503153.

    • Search Google Scholar
    • Export Citation
  • 29.

    Khan MG, Alam MS, Podder MP, Itoh M, Jamil KM, Haque R, Wagatsuma Y, 2010. Evaluation of rK-39 strip test using urine for diagnosis of visceral leishmaniasis in an endemic area in Bangladesh. Parasit Vectors 3: 114.

    • Search Google Scholar
    • Export Citation
  • 30.

    Singh D, Pandey K, Das VN, Das S, Kumar S, Topno RK, Das P, 2009. Novel noninvasive method for diagnosis of visceral leishmaniasis by rK39 testing of sputum samples. J Clin Microbiol 47: 26842685.

    • Search Google Scholar
    • Export Citation
  • 31.

    Vaish M, Singh OP, Chakravarty J, Sundar S, 2012. rK39 antigen for the diagnosis of visceral leishmaniasis by using human saliva. Am J Trop Med Hyg 86: 598600.

    • Search Google Scholar
    • Export Citation
  • 32.

    Chakravarty J, Kumar S, Kumar R, Gautam S, Rai M, Sundar S, 2011. Evaluation of rk39 immunochromatographic test with urine for diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg 105: 537539.

    • Search Google Scholar
    • Export Citation
  • 33.

    Cruz I, Chicharro C, Nieto J, Bailo B, Cañavate C, Figueras MC, Alvar J, 2006. Comparison of new diagnostic tools for management of pediatric Mediterranean visceral leishmaniasis. J Clin Microbiol 44: 23432347.

    • Search Google Scholar
    • Export Citation
  • 34.

    Chulay JD, Bryceson AD, 1983. Quantitation of amastigotes of Leishmania donovani in smears of splenic aspirates from patients with visceral leishmaniasis. Am J Trop Med Hyg 32: 475479.

    • Search Google Scholar
    • Export Citation
  • 35.

    Badaró R, Benson D, Eulálio MC, Freire M, Cunha S, Netto EM, Pedral-Sampaio D, Madureira C, Burns JM, Houghton RL, David JR, Reed SG, 1996. rK39: a cloned antigen of Leishmania chagasi that predicts active visceral leishmaniasis. J Infect Dis 173: 758761.

    • Search Google Scholar
    • Export Citation
  • 36.

    Thakur CP, Ahmed S, 2001. Observations on amphotericin B treatment of kala-azar given in a rural set up in Bihar, India. Indian J Med Res 113: 1418.

    • Search Google Scholar
    • Export Citation
  • 37.

    Sundar S, Singh A, Rai M, Prajapati VK, Singh AK, Ostyn B, Boelaert M, Dujardin JC, Chakravarty J, 2012. Efficacy of miltefosine in the treatment of visceral leishmaniasis in India after a decade of use. Clin Infect Dis 55: 543550.

    • Search Google Scholar
    • Export Citation
  • 38.

    Solano-Gallego L, Rodríguez A, Iniesta L, Arboix M, Portús M, Alberola J, 2003. Detection of anti-Leishmania immunoglobulin G antibodies in urine specimens of dogs with leishmaniasis. Clin Diagn Lab Immunol 10: 849855.

    • Search Google Scholar
    • Export Citation
  • 39.

    Brandtzaeg P, Fjellanger I, Gjeruldsen ST, 1970. Human secretory immunoglobulins. I. Salivary secretions from individuals with normal or low levels of serum immunoglobulins. Scand J Haematol Suppl 12: 383.

    • Search Google Scholar
    • Export Citation
  • 40.

    Roitt I, Lehner T, 1983. Immunology of Oral Diseases, 2nd edition. Oxford: Blackwell Scientific Publications, 279280.

  • 41.

    Ghatei MA, Hatam GR, Hossini MH, Sarkari B, 2009. Performance of latex agglutination test (KAtex) in diagnosis of visceral leishmaniasis in Iran. Iran J Immunol 6: 202207.

    • Search Google Scholar
    • Export Citation
  • 42.

    Boelaert M, El-Safi S, Hailu A, Mukhtar M, Rijal S, Sundar S, Wasunna M, Aseffa A, Mbui J, Menten J, Desjeux P, Peeling RW, 2008. Diagnostic tests for kala-azar: a multi-centre study of the freeze-dried DAT, rK39 strip test and KAtex in East Africa and the Indian subcontinent. Trans R Soc Trop Med Hyg 102: 3240.

    • Search Google Scholar
    • Export Citation
  • 43.

    Carvalho SF, Lemos EM, Corey R, Dietze R, 2003. Performance of recombinant K39 antigen in the diagnosis of Brazilian visceral leishmaniasis. Am J Trop Med Hyg 68: 321324.

    • Search Google Scholar
    • Export Citation
  • 44.

    Hatam GR, Ghatee MA, Hossini SM, Sarkari B, 2009. Improvement of the newly developed latex agglutination test (Katex) for diagnosis of visceral lieshmaniasis. J Clin Lab Anal 23: 202205.

    • Search Google Scholar
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 16 16 7
Full Text Views 365 90 0
PDF Downloads 81 14 0
 
Membership Banner
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save