Mutational analysis of the HEV isolates from patients with acute liver failure in north India

Abstract

S 21 ANNUAL CONFERENCE—2013 A cu te Lver Fa ilu re admission to predict complications and survival among patients with ALF. Prognostic value of serum actin free Gc-globulin levels were also compared with the Kings College Hospital criteria in predicting mortality. Methods: two consecutive ALF patients admitted from October 2011 to December 2012 were followed up until death or complete recovery. All had arterial ammonia and serum actin free Gc-globulin estimated at admission. Logistic regression analysis was performed to identify independent predictors of mortality. Receiver operating characteristic curve analysis was also done. Results: Twenty five (78.1%) patients died. Non-survivors had significantly lower mean serum actin free Gc-globulin levels than survivors (87.32 v 180mg/L; P <0.001) and higher arterial ammonia levels than survivors (128 v 90.57mmol/l; P <0.05). A receiver operating characteristic curve analysis showed that serum actin free Gc-globulin level cut-off of #124 mg/L was found to predict mortality with 92% sensitivity and 71.4% specificity. The corresponding figures for the Kings College Hospital criteria were 60% and 85%. Receiver operating characteristic curve analysis comparing arterial ammonia and serum actin free Gc-globulin levels in predicting mortality revealed an area under curve (AUC) of 0.771 for serum actin free Gc-globulin compared with an AUC of 0.737 for arterial ammonia .Patients with lower serum actin free Gc-globulin levels also developed more complications, including sepsis (P <0.001), Acute Kidney Injury (P <0.05), and deeper encephalopathy (p NS). Logistic regression analysis showed that serum actin free Gc-globulin and arterial ammonia at admission were independent predictors of mortality (odds ratios 0.982 and 1.024 respectively). Incorporating these variables, a score predicting mortality risk at admission was derived: 0.987 + 0.024arterial ammonia level (mmol/l) -0.018 serum actin free Gc-globulin levels (mg/L). Conclusion: Serum actin free Gc-globulin level at presentation is predictive of outcome and can be used for risk stratification. It is a better mortality indicator compared to arterial ammonia levels. Corresponding author. Premashish Kar. E-mail: premashishkar@gmail.com MUTATIONAL ANALYSIS OF THE HEV ISOLATES FROM PATIENTS WITH ACUTE LIVER FAILURE IN NORTH INDIA Jayanta Borkakoti, Giasuddin Ahmed, Rajib Kishore Hazam, Premashis Kar PCR Hepatitis Laboratory, Department of Medicine Maulana Azad Medical College and associated Lok Nayak Jai Prakash Hospital, University of Delhi, New Delhi, India, Department of Biotechnology, Gauhati University, Assam, India S16 Background and Aims: Previous studies suggest that the genotype of Hepatitis E virus influences the severity of hepatitis E. Viral capsid proteins play extensive role in interacting cellular proteins during capsid assembly and virus entry. Studies at the molecular level of the capsid protein of the HEV genome are hence warranted. We aimed to scrutinize the molecular alterations in the HEV genome in patients with acute liver failure and acute viral hepatitis. Methods: A total of 28 patients with acute liver failure and 155 patients with acute viral hepatitis were screened for the study during the years 2010-2012. HEV IgM was detected by anti HEV IgM ELISA. HEV RNA was detected using Viral RNA extraction kit. The Open Reading Frame 2 (ORF 2) region of the HEV genome was amplified using Reverse Transcriptase PCR. Representative samples were directly sequenced. Full length nucleotide sequences of HEV isolates were retrieved from the Gen Bank /EMBL/ DDBJ databases and compared with the strains. Sequences were aligned by CLUSTAL W software. Results: The mean age of the AVH and ALF patients were 23.9 3.71 years and 24.6 3.33 years respectively. HEV RNA was detected in 84 (54.19%) AVH and 22 (78.57%) ALF patients respectively. A total of 15 nucleotide substitutions at various positions of the ORF 2 were observed after aligning the obtained sequence of ALF patient with the other reference sequences. The nucleotide substitutions obtained were mainly silent substitutions with some conserved substitutions with only one amino acid change from Lysine to Cysteine. Conclusion: The single amino acid substitution and the silent substitutions may be associated with the poor outcome in ALF patients. The underlying mechanisms for the involvement of silent substitutions in the HEV genome with the progression to acute liver failure remains unknown and needs to be clarified in large number of cases. Corresponding author. Jayanta Borkakoti. E-mail: borkakoti.jayanta@gmail.com MATERNAL NUTRITIONAL PARAMETERS AS PREDICTIVE MARKER FOR ACUTE LIVER FAILURE IN HEPATITIS E INFECTION DURING PREGNANCY Sheetal Sharma, Ashok Kumar, Premashish Kar, Sarita Aggarwal Department of Obstetrics &Gynecology,Maulana AzadMedical College and associated Lok Nayak Hospital, New Delhi, India, Department of Medicine, Maulana Azad Medical College and associated Lok Nayak Hospital, New Delhi, India, Department of Biochemistry, Maulana Azad Medical College and associated Lok Nayak Hospital, New Delhi, India Background: Hepatitis E virus (HEV) infection in pregnant women is more common and fatal during the third trimester. Pregnant women in developing countries suffer