Multiplex polymerase chain reaction-based serotype analysis of dengue virus during 2015 dengue outbreak in Pakistan

Dengue is one of the most important arthropod-borne viral diseases. It is endemic in > 125 countries including Pakistan, with a global incidence of 50-200 million. We determined the frequency of different serotypes of dengue virus to highlight its hyperendemicity in Rawalpindi, Pakistan. Between May and October 2015 we analysed the serum samples of 140 patients with a suspicion of dengue, using ELISA and multiplex polymerase chain reaction. One hundred and eight were infected with serotype 2, 16 with serotype 3, 7 with serotype 4 and 3 with serotype 1. Three patients were infected with serotypes 1 and 2, and 1 each with serotypes 1 and 4 and serotypes 2 and 3. Incidence of dengue has increased many fold in the past 50 years and has expanded to areas that were previously free from the disease. Serotype 2 was predominant in our population followed by serotype 3. There is currently no specific treatment for dengue, and vector control and vaccination are the only effective methods to prevent future outbreaks

Outbreak of chickenpox in a military center in northern Pakistan

Objective: To study the outbreak of chickenpox in a military training center in Northern Pakistan

Study Design: Cross sectional study

Place and Duration of Study: Combined Military Hospital [CMH] Attock from 1st Nov 2007 to 30th Jan 2008

Material and Methods: Patients with clinical suspicion of chickenpox during the outbreak were confirmed by laboratory tests including complement fixation test using commercial antigen of Virion/Serion of Germany and Immunoglobulin M for Varicella zoster virus by enzyme linked immunosorbent assay of Vircell, Spain. Moreover, direct immunofluorescence using BioRad kit was also done in patients with active lesions

Results: A total of 102 cases occurred in the present outbreak. The age of the patients ranged from 16 to 35 years with a mean of 21.16 + 4.47 years. From a total of 102 patients, 60 [58.8%] were positive for antibodies against Varicella zoster virus by complement fixation test, 30 [29.4%] were positive by ELISA and 12 [11.7%] were positive on Immunofluorescence testing. None of the admitted cases developed any complication and their hospital stay remained uneventful

Conclusion: This outbreak of chickenpox in a military training center affected a considerable number of military recruits, led to a number of hospital admissions and had an adverse effect on training activities. There is a need to formulate a policy to offer vaccination against Varicella zoster virus to all unimmunized individuals before induction in the military to avoid unnecessary interruption in training and academic activities

Seroprevalence of transfusion transmissible infections in blood donors: a three year experience

Objective: To determine seroprevalence of transfusion transmissible infections [TTIs] in blood donors and to compare the present results with the data from studies conducted in past

Study Design: Descriptive study

Place and Duration of Study: Armed Forces Institute of Transfusion [AFIT], Rawalpindi from January 2010 to December 2012

Material and Methods: All the blood donors who had donated blood at AFIT during the three year study period were included. Prior to blood donation at the institute, all the donors were subjected to a preset, structured questionnaire to determine their eligibility for donation as per the criteria set by the institute. Donors7 serum samples were screened for hepatitis B virus [HBV], hepatitis C virus [HCV], human immunodeficiency virus [HIV] and syphilis by using HBsAg, anti-HCV, HIV antigen-antibody combination assay and syphilis antibody test, respectively. The repeatedly reactive samples were considered as true reactive

Results: A total 160552 blood donors were screened during the study period. The mean age of the donors was 29 ++/- 10.2 years [Range: 18 - 60 years]. Out of these, 158144 [98.5%] were male donors and 2408 [1.5%] were female donors. 7385 [4.6%] donors were volunteer and 153167 [95.4%] donors the replacement donors. The seroprevalence of TTIs in the donors for HBV, HCV, HIV and syphilis was 2385 [1.48%], 4194 [2.61%] 26 [0.02%] and 1520 [0.95%], respectively. The seroprevalence of HBV was higher and statistically significant [p value<0.05] in Gp-II [31-45 years] and the seroprevalence of both HCV and Syphilis was higher and statistically significant [p value <0.05] in both GP-II [31-45 years] and GP-III [46 years and above] when compared with overall seroprevalence of the respective infections in all age groups

Conclusion: This study highlights that the seroprevalence of HBV and HCV is decreasing in our blood donors, but still it is an important risk factor for spread of these infections. The seroprevalence of HIV is rising gradually in the blood donors

End of treatment response in patients treated for hepatitis c virus with standard interferon and ribavirin based on viral load

To determine the End-of-Treatment-Response [ETR] to standard interferon and ribavirin based regimen in patients of chronic hepatitis C and to compare the ETR response in low and high viral load groups. Descriptive study. Virology Department, Armed Forces Institute of Pathology [AFIP], Rawalpindi, from March 2012 to May 2013. Patients with chronic hepatitis C virus infection were included in the study. Pre-treatment viral load was determined by RoboGene Quantification kit. Based on viral load, the 400 patients were divided into two equal groups of low viral load [< 800,000 IU/ml] and high viral load [> 800,000 IU/ml]. The patients were treated with standard interferon alpha [3 million units subcutaneously thrice weekly] and ribavirin [10.6 mg/kg body weight] for 6 months. ETR was measured using Sacace Biotechnologies Qualitative kit. Chi-square test was used to compare the ETR in the two viral load groups. P-value < 0.05 was considered as significant. Out of 400 patients, 206 [51.5%] were males and 194 [48.5%] were females. Two hundred seventy [67.5%] patients achieved ETR and 130 [35.5%] failed to do so. In low viral load group, 145 [72.5%] patients achieved and 55 [27.5%] patients did not achieve ETR. In high viral load group, 123 [61.5%] patients achieved and 77 [38.5%] did not achieve ETR. The difference in ETR between low and high viral load groups was statistically significant [p=0.019]. End-of-treatment-response in patients treated for hepatitis C virus with standard interferon and ribavirin was greater in patients with low viral load as compared to patients with high viral load

Haematological manifestations and frequency of FAB subtypes in patients of acute meyoloid leukaemia: a single center study

To determine the clinical/haematological manifestations and frequency of different subtypes of Acute Myeloid Leukaemia [AML] according to the French-American-British [FAB] classification. Descriptive study. The study was carried out at haematology department of Armed Forces Institute of Pathology [AFIP], Rawalpindi from January 2011 to September 2012. Retrospective review of documents of patient diagnosed to have acute myeloid leukaemia on bone marrow aspiration was done. Patient's age, gender, major signs and symptoms at time of presentation and haematological parameters of peripheral blood and bone marrow were noted. The subtype of AML according to FAB classification was also documented. Data was entered and analyzed in SPSS 16.0. During the selected study duration acute myeloid leukaemia was diagnosed in 173 patients on bone marrow examination. Out of these 123 [71.1%] were males and 50 [28.9%] were females. Thirty [17.3%] of the patients fell in paediatric age group [< 15 years] while the remaining 143 [82.7%] were in adult age category [> 15 years]. The mean age of presentation was 9 years among paediatric patients and 44.5 years among adults. The overall mean age of both these two groups was 38.4 years [3-84 years]. Fever [71.6%], generalized weakness [34.1%] and pallor [23.7%] were the three main complaints of the patients, followed by splenomegaly and lymphadenopathy. The mean total leukocyte count, haemoglobin and platelet count were 57.4 x 10[9]/L, 7.9 g/dL and 54 x 10[9]/L respectively. AML-M[2] was found to be the most frequent FAB AML subtype among 72 [41.6%] paediatric and adult patients. The main signs and symptoms of the patients of AML presenting to our centre were fever, generalized weakness and pallor. AML-M[2] was found to be the most common FAB subtype among AML in children and adults

Molecular and serological assessment of chronic HBV carriers and additional burden of applying updated guidelines in Pakistan

To assess the additional burden of the patients eligible for treatment, based on recommendations on viral load, in the light of 2009 version of AASLD guidelines, as compared to 2004 guidelines and to determine the frequency of HBeAg in chronic HBV carriers. Descriptive cross-sectional study. Virology Department, Armed Forces Institute of Pathology, Rawalpindi, from November 2010 to January 2012. Persons with chronic HBV infection, reporting for HBV DNA PCR test, were included in the study and blood samples were collected. HBV DNA load was determined by Real Time PCR. HBsAg and HBeAg were tested by ELISA. Out of the 801 subjects positive for HBsAg, 74 [9.24%] were positive for HBeAg. Out of them, 113 [14.1%] had HBV DNA load > 100,000 copies/ml and were eligible for treatment according to AASLD 2004 guidelines. Forty one [5.1%] had HBV load between 10,000 and 100,000 copies/ml, and were additionally eligible for treatment as per AASLD 2009 guidelines. The 5.1% of 4.5 million estimated HBV carries in Pakistan comes to 229500. There was a low HBeAg positivity and HBV DNA positivity in our chronic HBV infected persons. Moreover, there is an increase of 229500 potential candidates for HBV treatment in Pakistan based on viral load testing, according to the AASLD 2009 guidelines when compared with 2004 guidelines. The increase in the number of candidates for treatment may require an additional expenditure of tens of billions of rupees

Frequency of abo and RH [D] blood groups among blood donors in Rawalpindi/Islamabad area

To determine the frequency of various ABO and Rh [D] blood groups among the blood donors in Rawalpindi/Islamabad area of Pakistan. Cross sectional descriptive study. Armed Forces Institute of Transfusion [AFIT], Rawalpindi from Sep 2010 to Jan 2011. A total of 4642 healthy adult, blood donors [volunteer and directed both] belonging to Rawalpindi/Islamabad region were included. From each donor 2 ml of EDTA blood was taken. ABO and Rh [D] blood grouping were carried out by tube method using commercially prepared antisera. The frequency of each type was calculated. Among ABO blood groups the most frequent was B with 1593 subjects [34.3%], followed by O with 1454 [31.3%], A with 1124 [24.2%], and AB with 471 [10.1%] individuals. The Rh [D] positive were 4226 subjects [91%] and Rh [D] negative were 416 [9%]. Among Rh [D] positive cases, blood group B was most frequent with 1476 [34.9%] individuals, however among Rh [D] negative individuals the most frequent blood group was O with 149 individuals [35.8%]. 'B' was most frequent among the ABO blood groups. Knowledge of prevalence of various blood groups in the area will help in managing the transfusion services

Prevalence of hepatitis C virus ribonucleic acid in antibody screened blood in Northern Pakistan

Blood donors are currently screened for antibodies to hepatitis C virus [HCV] by enzyme immunoassay. This pilot study aimed to estimate the prevalence of viremic and potentially infectious donors by the HCV-RNA polymerase chain reaction [PCR] in donor population of Northern Pakistan and also to examine the prospects of adding nucleic acid testing [NAT] to serological screening protocols for donated blood with the purpose of reducing the risk of transfusion-transmission of HCV infection. Descriptive cross sectional study. The study was conducted at Armed Forces Institute of Transfusion, Rawalpindi in January 2010. Fifteen hundred samples negative for anti-HCV [EIA, third generation] were screened for HCV RNA using RT-PCR method for qualitative detection of HCV RNA, with a lower limit of detection of 200 Ill/ml. The prevalence of HCV RNA positivity in HCV antibody negative infectious blood could not be estimated as all blood donations were negative for HCV RNA. The prevalence of HCV transmission in anti-HCV screened blood has not yet been documented in Pakistan. The sample size [1500] of the pilot study was not large enough to predict prevalence of HCV RNA in anti-HCV tested blood donors. The introduction of NAT to screen donated blood has shown to improve blood -safety in developed world and is likely to have greater impact in Pakistan because of high prevalence of HCV in general population

Hepatitis A virus infection - shifting epidemiology

To determine the age distribution in HAV infection and seasonal variations in the prevalence of acute viral hepatitis caused by hepatitis A virus. A descriptive study. The study was carried out on the patients reporting at Virology Department, Armed Forces Institute of Pathology [AFIP], Rawalpindi, for determination of hepatitis A virus [HAV] IgM antibody, from July 2003 to June 2004. Altogether 626 patients with clinical suspicion of hepatitis A virus infection were referred to AFIP Rawalpindi for this test. Blood samples were collected and sera were separated and transferred to plastic aliquots that were stored at -20oC in a retrievable fashion until utilized in testing. The testing for ant-HAV IgM was carried out with the help of a commercial Enzyme Linked Immunosorbant Assay [ELISA] using reagent kits of DiasSorin [Germany] for HAV IgM antibodies. The HAV IgM positive rate was 40.57% [252/626]. Those tested included the sporadic cases as well as the patients from outbreak in two schools of Nowshera cantonment. The age of patients testing positive for HAV IgM, ranged from 03 to 27 years. There was a statistically significant seasonal difference in rate of positivity in different months of the calendar year. An outbreak of HAV infection was seen in the children of two neighboring schools of a cantonment, in which 44 children in different classes developed clinical jaundice. HAV infection occurs in a significant proportion of young people with a clinical suspicion of HAV infection. There is a changing trend of developing hepatitis A in the age beyond 18 years and in outbreaks, which was not there in our patients previously due to universal immunity found against HAV by the age of 18. It was because of chances of consumption of polluted food

Outbreak of dengue fever in Mangla and Mirpur area

Dengue virus infection has been endemic in Pakistan. An outbreak of febrile illness associated with thrombocytopenia was reported from Mangla and adjoining areas in vicinity of Mangla Dam during July-August 2003. We carried out a serological investigation of the outbreak by serosurveillance. Sera from 52 representative cases were received for viral studies at our institute. Aim of the study was to rule out dengue infection in these cases. Descriptive laboratory based. Department of Virology, Armed Forces Institute of Pathology, Rawalpindi. According to clinical history all the, patients had fever of 3-10 days duration associated with thrombocytopenia. Suspecting Dengue infection, their acute sera were tested for Dengue IgM, by the kit manufactured by Diagnostic Automation Inc., Dengue virus IgM ELISA USA. Among 52 single sera from these cases. Dengue IgM antibodies were detected in 38[73%] cases. Among the sero-positive cases, 7[18.4%] had Vivax malaria. 32[84%] were males with mean age of 34 years and 6[16%] were females with mean age of 27 years. Platelets were below 150,000/micro l in all the cases. Occurrence of Dengue fever in northern Pakistan has been documented and this should now be considered in the differential diagnosis of undiagnosed cases of fever, moreover concomitant malaria infection emphasizes the need for sustainable, community-based mosquito control

Categorization of hbv carriers

Hepatitis B virus [HBV] carrier state can broadly be classified into ordinary and super categories on the basis of Hepatitis B 'e' Antigen [HBeAg] and antibody to Hepatitis B 'e' antigen [anti HBe]. A total of 523 cases of HBV infection were included in this retrospective study for the purpose of subcategorization. Out of 433, males 73.44% were ordinary carriers [HBeAg Non-reactive but having antibody to that antigen], 18.01% super carriers [converse serological state] and 8.41% doubly negative carriers. In 90 females tested, 65.55% were ordinary carriers, 26.66% were super carriers and 7.77% were doubly negative. This pattern may help clinicians in prophylaxis to their contacts and for prognostic considerations

Hepatitis B in health care personnel

The exposure of health care personnel to blood, its products and carriers of Hepatitis B is much greater as compared to other segments of population. This study gives prevalence of past exposure and carrier state of HBV in health care personnel of our setup. It has been seen in our study that the nurses and laboratory workers are the worst affected. Two hundred and thirty one health care personnel [155 laboratory workers, 28 members of dental staff and 48 operation theatre staff] were tested for their ongoing HBV carrier state [HBsAg] and evidence for exposure to HBV in the past.Out of these 92 [39.8%] showed evidence of exposure to the virus in the past by being antiHBc positive.Out of the exposed persons 18 [7.7% of the total] were the HBsAg carriers and 61 [26.4%] had already developed immunity against the HBV by virtue of being anti-HBs positive