scholarly journals Detection of LD body in Peripheral Blood Buffy-coat from Suspected Kala-azar Cases of Bangladesh

2010 ◽  
Vol 3 (2) ◽  
pp. 27-32 ◽  
Author(s):  
Chandra Kumar Roy ◽  
Sofia Andalib Sariullah ◽  
Ahmed Abu Saleh ◽  
Md Ruhul Amin Miah
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Peripheral Blood ◽  
Venous Blood ◽  
Buffy Coat ◽  
P Value ◽  
Smear Microscopy ◽  
College Hospital ◽  
Kala Azar ◽  
Diagnostic Sensitivity And Specificity

The present study has been carried out in an attempt to observe the usefulness of peripheral blood buffy-coat microscopic examination for detection of LD-body from clinically suspected Kala-azar cases. Total 127 individuals are included in this study as cases and controls. Among them 67 are clinically suspected Kala-azar cases included from one of the Kala-azar endemic areas of Bangladesh and 60 are taken as endemic and nonendemic healthy controls. Peripheral venous blood and bone marrow aspirate collected and tested for LD bodies from clinically suspected Kala-azar cases in Rajshahi Medical College Hospital, during the period of July 2006 to June 2007. Serum from all cases and controls were tested by rK39 ICT for serological identification of visceral leishmaniasis. Laboratory examinations were performed in the Department of Microbiology and Immunology, BSMMU, Dhaka. Out of 67 study cases bone marrow smear microscopy for LD bodies positive in 44 (66.67%) and buffy-coat smear microscopy positive for 21 (31.34%); diagnostic sensitivity and specificity of buffy-coat smear microscopy was 47.72% and 100% in comparison with bone marrow examinations. Buffy-coat smear made form the 23 splenomegalic patients, microscopy revealed significant number 17 (73.91%) of the cases were positive for LD-body and all of the four hepato-splenomegalic patients were positive (p-value >0.05, reached from 2 test). Results of rK39 ICT for detection of visceral leishmaniasis cases show 100% sensitivity and 90% specificity. Peripheral blood buffy coat examination for LD body in splenomegalic cases of Kala-azar may replace bone marrow examinations.Bangladesh J Med Microbiol 2010; 03 (02): 27-32

scholarly journals Evaluation of qPCR on blood and skin microbiopsies, peripheral blood buffy coat smear, and urine antigen ELISA for diagnosis and test of cure for visceral leishmaniasis in HIV-coinfected patients in India: a prospective cohort study

BMJ Open ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. e042519
Author(s):  
Sophie I Owen ◽  
Sakib Burza ◽  
Shiril Kumar ◽  
Neena Verma ◽  
Raman Mahajan ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Hiv Infection ◽  
Peripheral Blood ◽  
Medical Science ◽  
Bone Marrow Aspiration ◽  
Buffy Coat ◽  
Tropical Medicine ◽  
Urine Antigen ◽  
Antigen Elisa

IntroductionHIV coinfection presents a challenge for diagnosis of visceral leishmaniasis (VL). Invasive splenic or bone marrow aspiration with microscopic visualisation of Leishmania parasites remains the gold standard for diagnosis of VL in HIV-coinfected patients. Furthermore, a test of cure by splenic or bone marrow aspiration is required as patients with VL-HIV infection are at a high risk of treatment failure. However, there remain financial, implementation and safety costs to these invasive techniques which severely limit their use under field conditions.Methods and analysisWe aim to evaluate blood and skin qPCR, peripheral blood buffy coat smear microscopy and urine antigen ELISA as non-invasive or minimally invasive alternatives for diagnosis and post-treatment test of cure for VL in HIV-coinfected patients in India, using a sample of 91 patients with parasitologically confirmed symptomatic VL-HIV infection.Ethics and disseminationEthical approval for this study has been granted by The Liverpool School of Tropical Medicine, The Institute of Tropical Medicine in Antwerp, the University of Antwerp and the Rajendra Memorial Research Institute of Medical Science in Patna. Any future publications will be published in open access journals.Trial registration numberCTRI/2019/03/017908.

scholarly journals QBC® for the diagnosis of human and canine american visceral leishmaniasis: preliminary data

2001 ◽  
Vol 34 (6) ◽  
pp. 577-581 ◽  
Author(s):  
Daniel B. Liarte ◽  
Ivete L. Mendonça ◽  
Francisco C.O. Luz ◽  
Elza A.S. de Abreu ◽  
Gustavo W.S. Mello ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Preliminary Data ◽  
Peripheral Blood ◽  
Buffy Coat ◽  
Promising Method ◽  
Blood Parasites ◽  
Leishmania Chagasi ◽  
Asymptomatic Carriers ◽  
Fluorescent Method

"Quantitative Buffy Coat" (QBC®) is a direct and fast fluorescent method used for the identification of blood parasites. Since Leishmania chagasi circulates in blood, we decided to test it in American visceral leishmaniasis (AVL). Bone marrow (BM) and peripheral blood (PB) of 49 persons and PB of 31 dogs were analyzed. QBC® was positive in BM of 11/11 patients with AVL and in 1/6 patients with other diseases. Amastigotes were identified in PB of 18/22 patients with AVL and in none without AVL. The test was positive in 30 out of the 31 seropositive dogs and in 28/28 dogs with Leishmania identified in other tissues. QBC® is a promising method for diagnosis of human AVL, and possibly for the exam of PB of patients with AVL/AIDS, for the control of the cure and for the identification of asymptomatic carriers. Because it is fast and easy to collect and execute, QBC® should be evaluated for programs of reservoir control.

scholarly journals Diagnosis of Kala-Azar by Nested PCR Based on Amplification of the Leishmania Mini-Exon Gene

1998 ◽  
Vol 36 (8) ◽  
pp. 2173-2177 ◽  
Author(s):  
Ken Katakura ◽  
Shin-Ichiro Kawazu ◽  
Toshimitsu Naya ◽  
Koichi Nagakura ◽  
Mamoru Ito ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Nested Pcr ◽  
Northwest China ◽  
Buffy Coat ◽  
Invasive Procedures ◽  
Blood Samples ◽  
Kala Azar ◽  
Pcr Method ◽  
Exon Gene

To diagnose visceral leishmaniasis (kala-azar), we have developed a nested PCR method based on amplification of the mini-exon gene, which is unique and tandomly repeated in the Leishmania genome. Nested PCR was sufficiently sensitive for the detection of DNA in an amount equivalent to a single Leishmania parasite or less. We examined the usefulness of this PCR method using bone marrow aspirates and buffy coat cells collected from kala-azar patients who had or had not received chemotherapy in northwest China. We obtained PCR positivity for all of the parasitologically positive bone marrow samples from the patients. Some ambiguities with the primary PCR results were eliminated by the subsequent nested PCR. The buffy coat samples from 7 of 12 patients with splenomegaly were positive by the nested PCR, although only 2 of them were positive for parasites by culture. However, buffy coat samples from nine children, whose splenomegaly has been reduced and clinically cured by antimony treatment, were all negative. Thus, this nested PCR method represents a new tool for the diagnosis of kala-azar with patient blood samples instead of bone marrow or spleen aspirates obtained by more invasive procedures.

scholarly journals Could kDNA-PCR in Peripheral Blood Replace the Examination of Bone Marrow for the Diagnosis of Visceral Leishmaniasis?

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Natalia Souza de Godoy ◽  
Marcos Luiz Alves Andrino ◽  
Regina Maia de Souza ◽  
Erika Gakiya ◽  
Valdir Sabbaga Amato ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Peripheral Blood ◽  
Gold Standard ◽  
Buffy Coat ◽  
Molecular Techniques ◽  
Parasitological Examination ◽  
Parasitological Diagnosis ◽  
Group 2 ◽  
Group 1

The aim of this study was to evaluate whether the molecular (kDNA-PCR) and parasitological diagnosis in peripheral blood (PB) could replace the invasive and painful bone marrow collection (BM) in the diagnosis of visceral leishmaniasis (VL). PB from suspected VL patients was evaluated by parasitological and molecular techniques using as the gold standard (GS) a combination of clinical, epidemiological, and immunochromatographic test (PB-rK39) results and parasitological examination of BM. Based on the GS, 38 samples from 32 patients were grouped: Group 1, 20 samples of VL cases, and Group 2, 18 samples of non-VL cases. In order to evaluate the parasitological and molecular techniques in PB, the samples were examined. From Group 1, PB kDNA-PCR was positive in 20 samples and in 19 of 20 in BM kDNA-PCR examination. However, the parasitological examination of buffy coat was insensitive, being able to detect only 4 cases from Group 1. All samples from Group 2 were negative. We concluded that, for the diagnosis of visceral leishmaniasis, the parasitological examination of peripheral blood was not useful; however, molecular diagnosis by kDNA-PCR, performed in peripheral blood, could be useful to replace the parasitological examination of bone marrow.

scholarly journals Clinical and Morphological Profile of Immune Thrombocytopenic Purpura in Children - A Five Year Study in a Paediatric Tertiary Health Care Centre of South India

2020 ◽  
Vol 7 (46) ◽  
pp. 2724-2729
Author(s):  
Ashida M. Krishnan ◽  
Deepthi Raj M.L ◽  
Priya V.S ◽  
Arya R.S
Keyword(s):  
Health Care ◽  
Bone Marrow ◽  
Immune Thrombocytopenic Purpura ◽  
Peripheral Blood ◽  
South India ◽  
P Value ◽  
Severe Thrombocytopenia ◽  
Care Centre ◽  
Health Care Centre ◽  
Thrombocytopenic Purpura

BACKGROUND Immune Thrombocytopenic Purpura (ITP) is one of the most commonly encountered disease in paediatric practice. Thorough clinical and morphological study of peripheral blood and bone marrow is required for confirming ITP. Clinicomorphological aspects of paediatric ITP is a less studied topic especially in developing countries like India. The objective was to study the clinical and morphological profile of paediatric cases of ITP. METHODS This is a 5-year record based retrospective study conducted in a paediatric tertiary health care centre in Kerala, South India. Data of all paediatric cases diagnosed as ITP including clinical presentation, clinical findings, blood counts, peripheral blood morphology, bone marrow morphology, and treatment response was collected and entered in SPSS software version 16.0 and analysed. For assessing correlation, chi-square test was used. RESULTS The age of children ranged from 3 months to 15 years. H/o viral fever was noted in 53 % cases. Cases which had moderate and severe thrombocytopenia were 74 % and 21 % respectively. Isolated thrombocytopenia was the most common peripheral blood picture observed with few cases showing coexisting eosinophilia and anaemia. All cases showed megakaryocyte proliferation in marrow with 9 % cases showing coexisting iron deficiency anaemia. Majority of cases showed rapid response to steroid / IVIG therapy and the response had no correlation with grade of thrombocytopenia (p value < 0.05). CONCLUSIONS Paediatric cases of ITP usually present following viral infections or vaccination, with worrisome bleeding episodes, petechiae, ecchymosis or purpura. KEYWORDS ITP, Paediatrics, Platelet Count, Thrombocytopenia, Vaccination

scholarly journals Bone Marrow and Peripheral Blood Cells Toxicity of a Single 2.0 Gy Cobalt60 Ionizing Radiation: An Animal Model

1970 ◽  
Vol 29 (2) ◽  
Author(s):  
Shittu Akeem ◽  
Olatunbosun Lukman ◽  
Khalil Eltahir ◽  
Olalere Fatai ◽  
Babatunde Abiola ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Marrow Cell ◽  
P Value ◽  
Peripheral Blood Cells ◽  
P Values ◽  
Cell Parameters ◽  
Post Irradiation ◽  
Total Body

BACKGROUND: Bone marrow is extremely vulnerable to damage caused by radiation therapy. Hence, bone marrow suppression is an important side effect of radiotherapy. Effective use of radiotherapy is therefore compromised by radiation-related injuries.MATERIAL AND METHODS: Six Guinea-pigs were recruited for the study of which three were subjected to total body irradiation with Co60 while the other three served as controls. Bone marrow and peripheral blood samples were collected before and at days 9, 14 and 21, post irradiation. Manual and automated counts were performed for bone marrow nucleated cells and peripheral blood cells respectively.RESULTS: Declining bone marrow cellularity was evident immediately post irradiation. Mean ± SD of marrow cell counted per mm3 were 121,924±281, 87,603±772, 121,367±375 and122,750±1000 pre-irradiation and days 9, 14 and 21, postirradiation (p-values 0.10, 0.27 and 0.29 respectively). Significant drops in counts were noticed on day 9 post-irradiation for all red cell parameters (p-values <0.05), for Total White Blood Cell Count and Neutrophil count (p-values <0.05) and also on days 14 and 21 for Lymphocytes (p-values <0.05) and on day 21 for Eosinophil/Basophil/Monocytes (p-value <0.05). A significant drop in platelets counts was also noticed on day 9 (p-value <0.05) which significantly increased above pre-irradiation value on day 21.CONCLUSION: Total body irrradiation with Co60 significantly affects the bone marrow with maximum reductions in marrow nucleated cells and peripheral blood cells counts on day 9 post irradiation. 

scholarly journals Comparison of KAtex, Bone Marrow Aspiration and DAT for the Diagnosis of Visceral Leishmaniasis

2019 ◽  
Vol 6 (1) ◽  
pp. 12-15
Author(s):  
Ishrat Sharmin ◽  
AKM Quamruzzaman ◽  
Rezina Parveen ◽  
M Abdulah Yusuf ◽  
Rashida Akter Khanam
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Bone Marrow Aspiration ◽  
Cross Sectional Study ◽  
Cross Sectional ◽  
Medical College ◽  
Kala Azar ◽  
History Of ◽  
One Year

Background: Newly developed KAtex test can be used as a non invasive tool for diagnosis of Kala-azar. Objectives: The aim of the present study was to compare KAtex, Bone marrow aspiration and DAT to diagnose VL. Methodology: This cross-sectional study was carried out in the Department of Microbiology at Dhaka Medical College, Dhaka, Bangladesh in collaboration with the Department of Parasitology, Institute of Epidemiology, Disease Control and Research (IEDCR), Dhaka, Bangladesh for a period of one year. Clinically suspected Kala-azar (VL) cases of different age and sex attending IEDCR, Dhaka from different Kala-azar endemic areas of Bangladesh were selected for this study. Patients having fever for more than 2 weeks, with or without splenomegaly, having history of loss of body weight following onset of fever were clinically suspected as Kala-azar cases. Microscopy and culture was performed in bone marrow (BM). KAtex was performed with urine sample. Agglutination of sensitized latex indicated presence of Leishmania donovani antigen in urine and thereby visceral leishmaniasis. No agglutination indicates absence of antigen in urine. DAT was done with serums of all cases. Result: Among 130 clinically suspected VL cases, 70 (53.85%) cases were BM positive and 60(46.15%) cases were BM negative. All the 70 BM positive cases were positive by KAtex and DAT. Among 60 BM negative cases, 15 were positive by KAtex and 23 were positive by DAT. The sensitivity of KAtex was 100.0% and specificity was 75.0%. The sensitivity of DAT was 100.0% and specificity is 61.6%. Conclusion: In conclusion, KAtex test is a good diagnostic tool for the detection of VL in comparison with DAT. Bangladesh Journal of Infectious Diseases, June 2019;6(1):12-15

The Immature Platelet Fraction in HIV Patients with Thrombocytopenia.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2095-2095
Author(s):  
Brian T. Garibaldi ◽  
Rupal B. Malani ◽  
Hsin-Chieh Yeh ◽  
Deborah Michell ◽  
Evan J. Lipson ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Bone Marrow ◽  
Platelet Count ◽  
Peripheral Blood ◽  
Reticulocyte Count ◽  
Control Group ◽  
P Value ◽  
Hiv Patients ◽  
Immature Platelet Fraction ◽  
The Mean

Abstract Thrombocytopenia is a common clinical feature of HIV infection. Given the number of possible etiologies of thrombocytopenia in a patient with known HIV, a peripheral blood test effective in determining the likely pathophysiologic basis of the thrombocytopenia would be a valuable clinical tool. Immature platelets are released early from the bone marrow in response to increased platelet turnover. These platelets contain residual megakaryocyte mRNA and have been termed reticulated platelets. A new assay, the Immature Platelet Fraction (IPF), measures the reticulated platelet count in peripheral blood. Patients with increased destruction of platelets from such conditions as ITP consistently have a higher IPF percent, while patients with decreased platelet production have a low or normal IPF percent. The goal of our study was to determine the performance characteristics of the IPF assay in HIV patients with thrombocytopenia and to see if the IPF percent could be used to help elucidate the etiology of low platelet counts in this group of patients. All adult patients admitted to the Johns Hopkins Hospital with a diagnosis of HIV and a platelet count less than 150,000 were eligible for enrollment. 62 patients were identified from February 2007 to June 2007. 34 control samples were obtained from inpatients with HIV who were not thrombocytopenic. In addition, 81 samples were available from non-HIV historical controls with normal platelet counts. The mean platelet count in the HIV thrombocytopenic group was 92,000 while the mean platelet count in the HIV control group was 254,000 (p value &lt;.001). The mean platelet count in the non-HIV historical control group was 274 (p=.34 when compared to the HIV control group). The mean IPF percent in the HIV thrombocytopenic group was 10.2% as compared to 6.8% in the HIV control group (p=.001). The mean IPF in historical non-HIV controls was 3.1% (p&lt;.001 for both the HIV thrombocytopenic and the HIV control group). Univariate analyses were conducted to identify potential individual predictors of a high IPF percent. Backward selection was then performed using multivariate linear models with a threshold Wald test p-value of 0.05. ITP, diabetes mellitus and cirrhosis were significantly associated with a higher IPF percent with a co-efficient (95% confidence interval) of 6.98 (3.05–10.91), 4.73 (1.39–8.06), and 14.18 (9.7–18.66), respectively. CD4 count, HIV viral load, hepatitis C and reticulocyte count were not correlated with IPF percent. Our results suggest that patients with HIV have increased platelet turnover as compared to patients without HIV. Thrombocytopenic patients with HIV have increased platelet turnover relative to both non-thrombocytopenic HIV patients and to historical non-HIV controls. History of ITP, diabetes mellitus, and cirrhosis are predictive of an elevated IPF percent. Reticulocyte count is not correlated to IPF percent, suggesting that a low reticulocyte count is not a reliable marker for decreased bone marrow production in HIV thrombocytopenia. It is unlikely that the IPF assay alone can be used to determine the pathophysiologic basis of thrombocytopenia in any single patient with HIV. Further work needs to be done to clarify the utility of the IPF assay in this group of patients.

scholarly journals Role of the Bone Marrow Examination among Undifferentiated Fever in Tropics

2018 ◽  
Vol 2 (2) ◽  
Keyword(s):  
Bone Marrow ◽  
Visceral Leishmaniasis ◽  
Bone Marrow Biopsy ◽  
Clinical Decision Making ◽  
Bone Marrow Aspiration ◽  
Clinical Decision ◽  
Bone Marrow Examination ◽  
Diagnostic Tools ◽  
P Value ◽  
Highly Sensitive

Background: Bone marrow aspiration and biopsy is one of the most important diagnostic tools for evaluation of undifferentiated fever. The positivity yield of these samples is highly specific that provides additional evidence for clinical decision making among the undifferentiated febrile cases. With this background we evaluated the bone marrow results of undifferentiated febrile cases for the last five years at B.P. Koirala Institute of Health Sciences, Dharan, Nepal. The objective of the study was to measure the sensitivity of the bone marrow investigations among undifferentiated febrile cohort. Methods: A retrospective study was performed from January 2010 till December 2014 evaluating bone marrow reports. Completed request forms and the histopathological reports of the bone marrow specimens were reviewed. Statistical data was analyzed using SPSS 17 and p-value of <0.05 was considered significant. Results: Over the half decade 319 specimens were collected for bone marrow biopsy out of that 27% were requested for undifferentiated fever. The mean and median age of the biopsy performed patients was 35 and 31 years respectively. Among all biopsy samples 59% was adequate for evaluation however among the undifferentiated febrile cases biopsy samples only 45% was adequate for evaluation. The sensitivity of bone marrow biopsy was 34%. There were 714 bone marrow aspiration samples of that 84% was adequate for evaluation. The most common etiological diagnosis for the undifferentiated fever from the marrow evaluation was visceral leishmaniasis (53%). The sensitivity of the bone marrow aspiration and aspiration or biopsy for visceral leishmaniasis was 95% and 98% respectively. (p value 0.03) Conclusion: Bone marrow aspiration is highly sensitive and specific for the diagnosis of visceral leishmaniasis among the undifferentiated fever at tropics in Nepal.

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