Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR

Srivastava, Pankaj; Mehrotra, Sanjana; Tiwary, Puja; Chakravarty, Jaya; Sundar, Shyam

doi:10.1371/journal.pone.0019304

Cited by 66 publications

(53 citation statements)

References 23 publications

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“…In addition, wellequipped laboratories are required for their performance. [13][14][15][16] Immunochromatographic nitrocellulose strips test (ICT) is simple and wildly used for rapid diagnosis of VL in human blood/serum; it is extremely sensitive and specific, and it is well-accepted at the field level. [17][18][19][20] Because of this acceptance, rK-39 ICT has been introduced in the KA elimination program that was started in 2005 by the governments of three countries (Bangladesh, India, and Nepal) with the aim to reduce the annual incidence of VL to less than 1 case per 10,000 people at the subdistrict level by 2015 or earlier.…”

Section: Introductionmentioning

confidence: 99%

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

Singh

Pandey

Das

et al. 2013

The American Society of Tropical Medicine and Hygiene

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Abstract. 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.

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Section: Introductionmentioning

confidence: 99%

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

Singh

Pandey

Das

et al. 2013

The American Society of Tropical Medicine and Hygiene

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“…Detection of Leishmania DNA in the peripheral blood has been previously reported [4][5][6][7][8][9][10] ; herein, we report the capacity to extract DNA from VL RDTs and use it as a molecular tool for leishmaniasis molecular epidemiology studies. We show that the proposed approach can be used as an RDT quality control tool and a tool to evaluate the geographic prevalence of VL subspecies, which is important information for clinical and public health use for monitoring Leishmania parasites prevalence, fluctuation, and transmission routes in time and space in a scaled-up manner.…”

Section: Introductionmentioning

confidence: 90%

A New Molecular Surveillance System for Leishmaniasis

Pandey¹,

Pandey²,

Mallik³

et al. 2014

The American Society of Tropical Medicine and Hygiene

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Abstract. Presently, global efforts are being made to control and eradicate the deadliest tropical diseases through the improvement of adequate interventions. A critical point for programs to succeed is the prompt and accurate diagnosis in endemic regions. Rapid diagnostic tests (RDTs) are being massively deployed and used to improve diagnosis in tropical countries. In the present report, we evaluated the hypothesis of, after use for diagnosis, the reuse of the Leishmania RDT kit as a DNA source, which can be used downstream as a molecular surveillance and/or quality control tool. As a proof of principle, a polymerase chain reaction-based method was used to detect Leishmania spp. minicircle kinetoplast DNA from leishmaniasis RDT kits. Our results show that Leishmania spp. DNA can be extracted from used RDTs and may constitute an important, reliable, and affordable tool to assist in future leishmaniasis molecular surveillance methods.

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“…Interestingly, VL was also found to promote the clinical progression of HIV disease and AIDS-defining conditions (3). Parasitological diagnosis remains the gold standard in VL diagnosis due to its high specificity and sensitivity; molecular diagnosis is mainly based on PCR assays (7). In contrast, diagnosis of HIV-VL coinfection is very challenging as the clinical manifestations typical of VL are not always present and/or the patients might demonstrate several nonspecific clinical signs.…”

mentioning

confidence: 99%

Specific Noninvasive Detection of Leishmania donovani in Desquamated Buccal Cell Swab Samples from Human Visceral Leishmaniasis-HIV Coinfected Patients

et al. 2014

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Diagnosis of human immunodeficiency virus (HIV) infection with visceral leishmaniasis (VL) coinfection is challenging.Specific diagnosis of VL in HIV-coinfected patients was evaluated by molecular methods in desquamated buccal swab samples, demonstrating 86.3% sensitivity and 98.3% specificity in controls. This test holds significant potential for development as a noninvasive diagnostic tool for VL in HIV-coinfected patients. Visceral leishmaniasis (VL) and human immunodeficiency virus (HIV) coinfection has emerged as a serious disease prototype and is on the rise in India and Central and South America, posing a difficult challenge to VL subjugation efforts (1, 2). Since its first report in 1985, 35 countries have reported cases of this coinfection (3). In India, the prevalence of HIV-VL coinfection had been reported to be 2% to 5% in a limited number of studies (4). HIV infection increases the risk of developing VL by 100 to 2,320 times in areas of endemicity (5, 6) and greatly increases the probability of relapse cases. Interestingly, VL was also found to promote the clinical progression of HIV disease and AIDS-defining conditions (3). Parasitological diagnosis remains the gold standard in VL diagnosis due to its high specificity and sensitivity; molecular diagnosis is mainly based on PCR assays (7). In contrast, diagnosis of HIV-VL coinfection is very challenging as the clinical manifestations typical of VL are not always present and/or the patients might demonstrate several nonspecific clinical signs. As tissue aspiration for demonstration of the amastigote stage is potentially associated with dangerous risks, diagnosis in field settings is often based on signs, symptoms, and serology. Specifically, splenomegaly is less frequent in coinfected patients (8,9), and 42% to 68% of these patients have other associated opportunistic infections with analogous symptoms, further complicating the clinical diagnosis of VL (10). Presentation of amastigotes in peripheral blood smears of HIV-VL coinfected patients has a positivity rate of only 50% to 53% (11,12). Moreover, as HIV-VL patients present a low level of immune response, the validity and usefulness of a rapid immunochromatographic test (ICT) using recombinant antigen k-39 (rk39) would be doubtful even in cases of HIV coinfection (32). Commonly used laboratory diagnostic procedures for VL involve analyses of the cellular and chemical constituents of blood. However, researchers argue that easily collectable sputum samples or buccal swabs would be more practical for VL diagnosis, especially under field conditions (13,14). Interestingly, several studies have reported the presence of amastigotes in extraordinary locations, such as the spinal fluid, lungs, tonsils, larynx, digestive tract, rectum, etc., in HIV-VL patients (3,15,16,17). Therefore, in the current study, we investigated the potential of the buccal swab as the sample for noninvasive, safe molecular detection of VL in HIV-VL patients on the basis of the presence of leishmanial genomic DNA (gDNA).The study p...

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Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR

Cited by 66 publications

References 23 publications

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

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

A New Molecular Surveillance System for Leishmaniasis

Specific Noninvasive Detection of Leishmania donovani in Desquamated Buccal Cell Swab Samples from Human Visceral Leishmaniasis-HIV Coinfected Patients

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