Evaluation of efficacy of serological methods for detection of HCV infection in blood donors: a single centre experience

Background and Objective: Blood transfusion is an essential and life-saving medical intervention. Despite multiple preventive measures transfusion-transmitted hepatitis C virus [HCV] infection continues to be a major healthcare issue in Pakistan. This study was conducted at National Institute of Blood Diseases and Bone Marrow Transplantation to evaluate the frequency of active HCV infection with or without co-infection in blood donors and also to determine comparative efficacy of Multisure HCV antibody assay [MHAA]; a new serological device

Methods: A total of 14652 blood donors visiting National Institute of Blood Diseases and Bone Marrow Transplantation [NIBD] Blood Bank from January 2013 to July 2014 were enrolled and screened for a range of blood borne infections such as HBV, HCV, HIV, malaria and syphilis. The HCV was screened simultaneously by Abbot Architect anti-HCV assay [CLIA] and MHAA. The active HCV infection was confirmed by nucleic acid testing [NAT] in reactive donors. Later; for determination of comparative efficacy of MHAA; all NAT positive samples were further tested using Monolis[TM], HCV blot 3.0, Anti-HCV plus V2 and Anti-HCV-MPBIO-EIA

Results: The HCV reactive sera were observed in 1.563 percent [226] donors. The NAT confirmed active HCV infection in 138 donors. Overall 27.84 percent of HCV positive donors exhibited co-infection either with HBV [2.57 percent], syphilis [22.78 percent]. Triple infection was not observed in any donor. The efficacy of MHAA is comparable to all the serological tests with a sensitivity of about 96.89 percent

Conclusion: Active HCV infection was present in 0.94 percent donors. With a sensitivity of 96.89 percent [95 percent CI: 95.66-98.12] the multi-parametric device MHAA can effectively detect HCV infection in donors. Thus, it can be used in limited health care settings for HCV screening

Seroprevalence of transfusion transmitted infections among different blood group donors at blood bank LUMHS, Hyderabad

Objectives: To study the prevalence of HBsAg, Anti-HCV, HIV, Syphilis and Malaria in blood donors

Methods: This is a cross sectional descriptive study, conducted at Blood bank and Transfusion center at Liaquat University of Medical and Health Sciences [LUMHS] Hyderabad, during the period from January, 2014 to June, 2015.A total of 4683 blood donors were screened for HBsAg, Anti-HCV and HIV on Architect 20001 [manufactured by Abbott], employing chemiluminescent microparticle immunoassay [CMIA]. For Syphilis, VDRL ICT kits were used and Malaria parasite was screen through MP slides. Blood grouping was performed by both forward and reverse methods

Results: This study showed a high frequency of HBsAg, VDRL and malaria positivity among the O-ve blood group donors, i.e. 3.70%, 9.25% and 0.61% respectively. Blood group B-ve individuals were commonly infected with HCV [12.5%] as compared with all other blood group donors. HIV is more commonly reported in A+ve blood group individuals. Blood group O+ve is more prevalent [37.41 %]

Conclusion: High frequency of HCV infection in blood donors advocates implementation of strict screening policy for donors and public awareness campaigns about preventive measures to reduce the spread of this infection as well as other transfusion transmissible infections

Diagnosis and phenotypic assessment of Pakistani Haemophilia B carriers

Objectives: 1: To assess the diagnostic utility of three polymorphisms [DdeI, XmnI and TaqI] and direct sequencing in haemophilia B [HB] carrier detection in Pakistani families. 2: To compare phenotypes of HB carriers with those of healthy females

Methods: The study was conducted from March 2014 till February 2016 at Khyber Medical University Peshawar and National Institute of Blood Diseases, Karachi. Individuals from HB families of Khyber Pakhtunkhwa [KP] and Federally Administered Tribal Areas [FATA] with known F9 mutation in the proband were enrolled into the study. FIX activity [FIX:C] levels were determined in all the participants. Bleeding scores [BS] and complete blood counts were performed in the female participants. Linkage analysis followed by targeted Sanger sequencing was carried out in all the study participants. Heterozygosity rate was determined for each polymorphism. Healthy females and the carrier groups were compared for bleeding phenotypes

Results: A total of 30 males and 48 females from 13 HB families were studied. The polymorphisms had a low heterozygosity rate. Direct sequencing determined the carrier status in all cases. The mean FIX:C was reduced whereas BS was raised in the carriers when compared with healthy females. A significant raise in white blood cells [WBCs] count was observed in the carriers

Conclusion: The three polymorphisms have a low heterozygosity rate in HB families from KP and FATA. Sanger sequencing is conclusive in determining carrier status in all the cases. FIX:C is low and BS is raised in the HB carriers in comparison to that of normal females. The mean WBCs count is significantly higher in the HB carriers than the normal females

Importance of immature platelet fraction as predictor of immune thrombocytopenic purpura

Background and Objective: Immune thrombocytopenic purpura [ITP] is a clinical syndrome in which a decreased number of circulating platelets [thrombocytopenia] manifests as a bleeding tendency, easy bruising [purpura] or extravasation of blood from capillaries into skin and mucous membranes [petechiae]. The diagnosis of ITP can be made clinically on the basis of symptoms, we need to see if ITP can be confirmed in patients by quantification of residual RNA containing immature platelets [megakaryocytic mass] or immature platelets fraction [IPF] using automated hematology analyzers [Sysmex XE-2100]

Methods: In order to check the efficacy of IPF% parameter of Sysmex XE-2100 a total of 231 patients of thrombocytopenia were included in this study. Complete blood count [CBC] was estimated. The data was statistically analyzed by SPSS version 17

Results: About 62 patients were diagnosed as ITP and 169 patients were diagnosed as non ITP on the basis of clinical history. The mean IPF % value of ITP patients was 16.39% and the IPF % value of Non ITP patients was 7.69% respectively. There was no significant difference in IPF% values with respect to time between sampling and acquisition of complete blood count. The diagnostic sensitivity of IPF% as biomarker for ITP and non-ITP was 85.71% [95% CI: 84.04% to 85.96%] and 41.76% [95% CI: 39.87% to 43.65%]

Conclusion: The mean IPF % value by Sysmex XE-2100 can be used to predict ITP

Evaluation of efficacy of various immunochromatographic rapid tests for dengue diagnosis

The immunochromatographic rapid tests facilitate the early diagnosis of dengue by providing evidence of the presence of virus specific proteins [antigens/ antibody] in human blood. Many products for rapid dengue diagnosis are available in the market; the performance of few selected products was evaluated and compared with enzyme linked immuno sorbent assays [ELISA]. Sera from a large number of patients [n=184] admitted to National Institute of Blood Diseases and Bone Marrow Transplantation [NIBD] were used to determine the efficiency of non-structural [NS] 1, IgA, IgG and IgM based rapid test devices for dengue diagnosis. The dengue NS1 antigen based device was least efficient while among the antibody based devices the dengue IgA rapid test [RDT] was comparatively better [specificity: 80.95%; sensitivity: 85.21%]. This device could detect both primary and secondary dengue infection and was found to be the most sensitive device at all point of sample collection. The dengue IgA RDT could be a cost effective and efficient rapid test device for timely dengue diagnosis at all levels of healthcare settings

Oxidative stress and total antioxidant status in acute leukemia at diagnosis and post remission induction phase

This study evaluated the activity of superoxide dismutase [SOD1], glutathione reductase [GR] and total antioxidant status [TAS] in the hemolysate and sera of patients with acute leukemia [AL] at diagnosis, post remission induction phase and in healthy controls. However, total antioxidant status and glutathione reductase activities normalized after remission induction phase in acute myeloid leukemia [AML] only whereas levels of SOD were reduced but not achieved the normal level in acute lymphoblastic leukemia [ALL]. TAS activity showed no difference in either sex among any subtype of acute leukemia but glutathione reductase level was significantly higher in female ALL patients. Activity of SOD was elevated in T-cell ALL and acute myelomonocytic leukemia however; no significant difference in the activity of GR and TAS was noted. Levels of antioxidant were reduced insignificantly in patients who achieved complete remission

Vitamin D levels in patients of acute leukemia before and after remission-induction therapy

To determine the levels of 25-hydroxyvitamin [25[OH]D3] in patients with acute leukemia and the effect of remission-induction chemotherapy. This study was case control, all newly diagnosed patients of acute leukemia between the age of one to sixty years and residents of Pakistan were enrolled and evaluated. Those who were unwilling or unable to provide written informed consent were excluded. All selected patients [n=86] were grouped in to acute myeloid leukemia [AML] and acute lymphoblastic leukemia [ALL]. AML was further categorized as A1 before remission-induction [n=17] and B1 after remission induction [n=13], ALL was further categorized as A2 before remission-induction [n=31] and B2 after remission induction [n=25]. The 25-hydroxyvitamin [25[OH]D3] levels were measured in the sera of all patients [before and after remission-induction] by one step delayed chemiluminescent micro particle immunoassay [CMIA].We compared 25[OH]D3 levels in all patients before and after the remission-induction chemotherapy. A total of 86 patients were analyzed, in which 60 patients were male. Mean age was 24.39 years [range, 1 to 60 years]; the mean levels of 25[OH]D in group A1 [n=17] was 17.70 +/- 3.2 ng/ml, in group B1 [n=13] 14.06 +/- 2.4 ng/ml, 19.07 +/- 7.08 ng/ml in group A2 [n=31], while 10.59 +/- 3.9 ng/ml found in group B2 [n=25]. 25[OH]D3 insufficiency was evident subnormal in majority of patients with acute leukemia and 25[OH]D3 were further reduced after remission-induction as compared to untreated group, difference was statistically significant when compared with each group

JAK2V617F mutation in chronic myeloid leukemia predicts early disease progression

To determine the association of JAK2V617F mutation along with BCR-ABL translocation or Philadelphia chromosome in chronic myeloid leukemia with early disease progression to advanced stages [accelerated phase or blast crisis] and poor outcome. Case series. National Institute of Blood Diseases and Bone Marrow Transplantation, Karachi, from February 2008 to August 2009. All the newly diagnosed cases of BCR-ABL or Philadelphia positive CML were tested for JAK2V617F mutation by Nested PCR. Demographic data, spleen size, hemoglobin levels, white blood cell and platelet counts were recorded. Independent sample t-test was used for age, haemoglobin level and spleen size. Fisher's exact test was applied to compare disease progression in JAK2V617F mutation positive and negative cases. Out of 45 newly diagnosed cases of CML, 40 were in chronic phase, 01 in accelerated phase and 04 in blast crisis. JAK2V617F mutation was detected in 12 [26.7%] patients; 09 [22.5%] in chronic phase, none in accelerated phase and 03 [75%] in blast crisis. During a mean follow-up of 8 months, 03 patients in chronic phase transformed in blast crisis and 02 into accelerated phase. Overall 08 out 0f 11 [73%] JAK2V617F positive patients either had advanced disease or showed disease progression. Only 2 of 20 [10%] available patients, negative for the mutation, showed disease progression by transforming into blast crisis [p < 0.001]. No statistically significant difference was seen in the age, spleen size, haemoglobin levels, white blood cells and platelets counts in JAK2V617F positive patients. JAK2V617F mutation was detected in 26.7% cases of chronic myeloid leukemia. A significant proportion of them showed early disease progression

Arteriovenous thrombosis in chronic renal failure patients receiving renal replacement therapy

To determine the frequency of thrombotic complications and to identify factors associated with arteriovenous thrombosis in patients of chronic renal failure receiving renal replacement therapy. A descriptive study. The study was carried out at Sindh Institute of Urology and Transplantation [SIUT], Karachi, from May 2003 to December 2003. Of the 3000 patients evaluated, 61 End Stage Renal Disease [ESRD] patients on regular dialysis, having recent renal transplant, were selected for the study after informed consent. These patients had arteriovenous thrombosis with temporary central lines thrombosis and vascular access problems. Cases of congenital or acquired thrombotic disorders, e.g. with malignancy, DIG, liver disease, systemic lupus erythematosus or other immunologic diseases, pregnancy or women using oral contraceptives, were excluded. Similarly, patients taking any type of anticoagulant therapy during the preceding one week were not included in the study. Findings were recorded in a structured questionnaire. Laboratory analysis was done after clinical and radiological evaluation. Thrombophilia screening included antithrombin, protein C, protein S deficiencies and lupus anticoagulant. Forty-seven out of 61 patients selected were positive for thrombophilia screening with protein C deficiency in 26.2%, protein S deficiency in 16.3%, antithrombin in 5%, lupus anticoagulant in 13.1% and combined deficiency was observed in 16.3%. Of the 3000 patients, 61 with frequency of 2% were found to be deficient in one or had combined deficiency of these. Thus, the study of ESRD patients presenting with arteriovenous thromboembolism emphasizes the need to reconsider the perception that this clinical entity is rare and requires further studies