Anti IgM Research Articles

Molecular Epidemiology of Hepatitis E Virus Genotypes in Blood Donors in Peshawar, Khyber Pakhtunkhwa

Background: Hepatitis E virus (HEV), an RNA virus, poses a significant global health threat, affecting 2.3 billion people annually, causing 70,000 deaths, over 50% of acute viral hepatitis cases. In Pakistan, HEV prevalence is high among pregnant women (26%), with a mortality rate of 26 to 30%. Blood transmission is a leading source of sporadic HEV infection.
 Methodology: The blood specimens were investigated with HEV to detect anti- IgM and IgG antibodies. Anti HEV IgM positive specimens confirmed the incidence of viral RNA by Polymerase chain reactions. The positive cases containing viral RNA were subjected to sequencing of ORF2.
 Result: A total of 3480 blood specimens were screened serologically by ELISA for the existence of anti HEV IgM and anti-HEV IgG antibodies. Of these 3480 samples, 170 (4.9%) were confirmed positive when screened for HEV antibodies among study population, there was a statistically significant difference observed between males and females (p=0.00001). From 170 positive samples, 60 (1.72 %) were positive for having anti-HEV IgM antibodies in plasma, whereas 110 (3.16 %) were found with anti-HEV IgG antibodies. Among 60 (1.72 %) IgM-positive samples, 18 (0.51 %) were confirmed positive by PCR for the presence of HEV-RNA. Genotype 1 was observed in all 18 HEV RNA-positive samples. 
 Conclusion: The findings of this study underscore the urgency of implementing HEV screening protocols before blood donation. The risk of transmission through blood products is evident, with 4.9% of screened blood specimens exhibiting HEV antibodies. Therefore, the National Blood Transfusion Regulatory Authority must incorporate HEV screening as a routine procedure in Pakistan.

Expression of IL32 and IL33 Genes in Tuberculosis Patients

In the present case-control study, blood specimens were obtained from (60) patients infected with TB from TB center in Baghdad city, and from (60) healthy persons as a control group during the period from January 2023 to December 2023. Results of demographic picture showed that the distribution of infections in males was 31(51.7%) compared to the control group 3(51.7%) and in females was 29 (48.3%) compared to the control group 29 (48.3%). The infection distribution according to age revealed that the highest infection rate was shown to be within the age group (<20 -29), followed by (30-39) then (40-50) years, which matched with the control group (<20 -29). According to residency, there were no significant differences between rural and urban residents. Mean ±Std anti TB IgM antibodies was (2.40±1.44) in comparison with controls (0.08±0.17), with highly significant difference (P<0.01). Also, Mean ±Std anti TB IgG antibodies was (1.42±0.59) in comparison to controls (0.11±0.21), with highly a significant differences P< 0.01. Mean±Std IL-33 was (20.38±6.53) in comparison to the controls (2.28±2.48), with highly significant differences P<0.01. Also, mean ±Std IL-32 was (10.61±2.24) in comparison to the control group (1.89±2.043) with highly significant differences P< 0.01. Expression of IL32 in TB patients showed positive reaction and it was highly affected in patients with TB compared to the control group, while expression of IL33 in TB patients showed positive reaction and was highly affected in patients with TB compared to the control group.

Distribution of Crimean-Congo Hemorrhagic Fever Virus (CCHF) Infections Among Animals and Human in Diyala Province

In this study, 30 blood samples were collected from Human patients infected with CCHF virus residing in Alrazi Hospital, 30 samples from cattles and 30 from goats from the veterinary medical clinic in Baquba city. Also, 15 blood samples were taken from healthy individuals, 15 samples from healthy cattles and 15 samples from healthy goats as control groups. The study was carried out during the period from, 15th January to 30th June 2023 in Diyala Province. According to the results, the (Mean±S.E) of Human anti IgM antibodies was 2.40±0.26 in patients with CCHF and 2.13±0.23 in cattles with non-significant differences, while the (Mean±S.E) of anti IgM antibodies was 1.72±0.15 in goats with a significant difference between goats and humans (p=0.03). The (Mean±S.E) of Human anti IgG antibodies was 1.42±0.11 in patients with CCHF and 1.27±0.12 in cattles with non-significant differences. Also, the mean ±S.E of anti IgG was 1.24±0.11 in goats with non-significant differences between the animals and Humans. The mean ±S.E of anti-IgM Human antibodies was 2.40±0.26 compared to the control group 0.08±0.05 with a highly significant difference p<0.001. The mean of anti-IgG human antibodies was 1.42±0.11 compared to the control group 0.13±0.06 with a highly significant difference p<0.001. The mean of anti-IgM cattle antibodies was 2.13±0.23 compared to the control group 0.08±0.04, with a highly significant difference p<0.001. The mean of anti-IgG antibodies was 1.27±0.12 compared to the control group 0.09±0.04 with a highly significant difference p<0.001. The mean of anti-IgM goat antibodies was 2.13±0.23 compared to the control group 0.08±0.04 with a highly significant difference p<0.001. finally, the mean of anti-IgG antibodies was 1.27±0.12 compared to the control group 0.09±0.04 with a highly significant difference p<0.001.

Effectiveness of Brucellosis on Polymorphisms of IL-1B and IL-10 Concentrations Among Iraqi Patients

In this study, (60) blood samples were taken from patients infected with brucellosis, and (60) samples were taken from healthy individual as a control group. The patients were visiting Emam Ali Hospital for Fever in Baghdad city, during the period from 1st February to 1st November 2023. The results of gender distribution showed that among patients infected with Brucellosis, 30 (50.0%) were males and 30 (50%) were females, while among the control group, 31(51.7%) were males and 29(48.3%) were females. Also the distribution of infections according residency were matched between urban and rural locations 30(50.0%) for both. While the distribution of Brucellosis among age groups showed that the highest incidence of infection was in the age groups (40-50) years and (17-29) years 22(36.7%) for each, followed by the age group (30-39) years 16(26.7%). The mean levels of anti Brucella antibodies IgM, IgG, IL-10, IL-1B in patients with brucellosis were (0.11±0.17), (0.09±0.15), (5.62±1.99), (4.92±1.76) respectively compared to their mean levels in the control group (2.01±1.17), (1.33±0.59), (60.68±26.81), (22.85±9.17) respectively with highly significant differences (p<0.01). The ROC test showed that the Sensitivity of Brucella IgM concentration was100% and Specificity 100% in Cut off >0.84. Also the Sensitivity of Brucella IgG concentration was 100% and Specificity 100% in Cut off >0.33. Also the Sensitivity of IL-10 concentration was 100% and Specificity 100% in Cut off >18.04, and the Sensitivity of IL-1B concentration was 100% and Specificity 100% in Cut off >9.08. There were mutations occurred with IL-1B gene ID 3553 in SNPs, rs1143627. The variation of wild TT was changed to CC, CC, TC, TC, CC, AG, CC, TC in 8 cases out of 11 in comparison with the control group, and the mutation occurred with IL-1B gene ID 3553 in SNPs, rs1143627. The variation of wild TT was changed to CC, CC, TC, TC, CC, AG, CC, TC in 8 cases out of 11 in comparison with the control group.

Hematological Manifestations of Antiphospholipid Antibodies: A Case Series from South India

Introduction: Antiphospholipid syndrome (APS) is known for its association with unexplained vascular thrombosis and pregnancy-related complications. However, APS can also present with various hematological abnormalities, including thrombocytopenia, autoimmune hemolytic anemia, Evan's syndrome, and thrombotic microangiopathy. While APS is often considered in patients with these manifestations, its laboratory testing yield and patterns in hematology clinics remain underexplored. In this study, we aimed to investigate the prevalence of laboratory positivity for APS in patients attending hematology clinics, even in the absence of meeting the clinical criteria for APS. Methodology: This retrospective study was conducted at two tertiary care centers in southern India from January 2018 to June 2023. Patients who visited hematology clinics and underwent APS workup were included. The APS workup involved evaluating Anti Cardiolipin IGG and IGM, Anti Beta2 Glycoprotein IGG and IGM, Lupus Anticoagulant, and ANA. The laboratory criteria for APS were based on the ACR/EULAR APS Criteria. Patients with transiently positive results were excluded from the study. Chi Square test was used for statistical analysis. Results: Among the 624 patients who underwent APS workup, 100 patients (16.03%) tested positive for at least one of the laboratory criteria for APS. Remarkably, a significant proportion (56%)of these patients did not fulfill the clinical criteria for APS. The most common hematological manifestations prompting APS testing in these 100 patients were thrombocytopenia (38%) and unprovoked thrombosis (35%). Additional hematological abnormalities included autoimmune hemolytic anemia (13%), prolonged activated partial thromboplastin time (APTT) (5%), Evan's syndrome (4%), pancytopenia (4%), and thrombotic thrombocytopenic purpura (TTP) (1%). Baseline characterstics of our study subjects are shown in Table 1 Anti-beta2 glycoprotein IGG or IGM antibodies were detected in 74% of APS-positive patients, and Anti-cardiolipin IGG or IGM antibodies were found in 57%. Lupus Anticoagulant (LA) was present in 59%. Additionally, 28% of the patients showed positivity for all three antibodies, indicating a significant overlap in laboratory findings. No statistically significant difference in antibody positivity patterns was observed between patients satisfying the clinical criteria for APS and those without, as shown in Table 2 Conclusion: Our study sheds light on the hematological presentations of APS in patients attending hematology clinics in southern India. Importantly, a substantial number(56%)of patients with APS lab test positivity did not meet the clinical criteria for APS. These findings emphasize the importance of considering APS as a potential diagnosis in patients presenting with hematological manifestations, especially thrombocytopenia and autoimmune hemolytic anemia. Identifying APS in such cases can guide appropriate therapeutic interventions and may help prevent pregnancy-related complications in affected females. Raising awareness among healthcare professionals about the diverse hematological presentations of APS is vital for improving patient outcomes and ensuring timely and accurate diagnoses

Status of anti-HEV IgG and IgM antibodies among the hemodialysis patients in southwest region of Iran.

Hepatitis E Virus (HEV) account for acute hepatitis, fulminant liver failure and chronic hepatitis worldwide. Several high risk groups including hemodialysis (HD) patients are at risk of HEV infection. Based on consequences of HEV infection it is important to determine the serological and molecular epidemiology of HEV in HD patients. The aim of this study was to evaluate the frequency of HEV antibodies and HEV RNA in HD patients. The sera of 84 HD patients were collected and tested for anti-HEV IgG and anti IgM antibodies using enzyme-linked immunosorbent assay (ELISA) at Golestan hospital in Ahvaz city during October 2014 and November 2014. HEV RNA was tested in HD patients using RT PCR. The prevalence of anti-HEV IgG was evaluated in the age group (52/84) > 50 and (31/84) < 50 years. Out of 84 patients, 52 (61.9%) were males and 32 (38.1%) females. The mean age of participants was 52 ± 1.57 years. 43/84 (51.19%) cases including 26/52 (50%) males and 17/32 (53.1%) females were positive for anti-HEV IgG (p=0.95). Among the 43 cases positive anti-HEV IgG 8 cases including 5 (9.61%) males and 3 (9.37%) females tested positive for anti-HEV IgM (p=0.729) while the HEV RNA was negative in HD patients. The distribution of anti-HEV IgG was 62.75% and 33.33% among the age group >50 and <50 respectively (p=0.015). This study showed high prevalence of anti-HEV IgG antibodies (51.19%) were observed among the HD patients while the HEV RNA tested negative in HD patients. The rate of HEV IgG is significantly higher with increased age. Further investigation require to identify the factors account for high seroprevalence of HEV in Ahvaz HD units.

S3177 The (Icteric) Eye of the Storm: A Case of Weil’s Syndrome

Introduction: Leptospirosis is one of the most prevalent zoonotic diseases affecting humans residing in tropical areas. We herein describe a case of a patient with leptospirosis acquired in the Bronx, NY. Case Description/Methods: A 63 year-old man with no significant medical history was evaluated for muscle pain, fatigue, and diarrhea of 3 days duration. He was feverish to 102 F and icteric. Laboratory evaluation revealed hemoglobin of 7.5 g/dL, total white count of 18 k/uL, platelet count of 40 k/uL, INR 1.5, creatinine of 5 mg/dL, urea nitrogen 85 mg/dL, creatine kinase 4000 U/L, alkaline phosphatase 192 U/L, total bilirubin 39 mg/dL, direct bilirubin > 23 mg/dL, AST 96 U/L, ALT 113 U/L and albumin 2.5 g/dL. Initial laboratory evaluation was negative for common and atypical infections. Radiologic evaluation revealed unremarkable non-contrast MRCP and US abdomen. Upon further detailed history, the patient reported that his basement was recently flooded by a devastating hurricane. Despite negative parasite blood smear and IgM antibodies for leptospira, empiric treatment for leptospirosis with Ceftriaxone resulted in resolution of all symptoms and lab abnormalities. Repeated blood work on follow-up visits at 4 weeks was noted with positive IgM antibodies for leptospira (Figure). Discussion: Leptospirosis is caused by a poorly staining gram-negative spirochetal bacteria. Transmission to humans occurs through mucous membranes, open skin wounds or after drinking, eating, swimming, or wading in water contaminated by stool or urine of infected rodents. Outbreaks have been documented in non tropical, urban areas affected by hurricanes, heavy rains or floods. In approximately 90% of cases, the disease is anicteric. Phase 1 includes viral-like illness, conjunctival suppuration followed by transient improvement. Phase 2 includes myalgia, nausea, vomiting, abdominal pain, hepatomegaly, increased serum bilirubin and serum aminotransferases. 10% of cases manifest as Weil’s syndrome, characterized by jaundice in the first phase, followed by second phase manifesting with high fever, profound jaundice with severely elevated serum bilirubin levels (up to 30mg/dL, mostly direct) and elevated serum transaminases. Renal and liver involvement are predominant and can lead to severe morbidity and death. Blood cultures, urine cultures and specific anti Leptospira IgM serology can assist with diagnosis. Antibiotic treatments, primarily Penicillin, 3rd generation Cephalosporins and Doxycycline, can benefit if administered at an early stage.Figure 1.: Laboratory evaluation during hospital stay and 4 weeks after discharge. Abbreviation: ALP, alkaline phosphatase; TB, total bilirubin; DB, direct bilirubin; AST, Aspartate transaminase; ALT, Alanine transaminase.

Development and application of a recombinant Envelope Domain III protein based indirect human IgM ELISA for Kyasanur forest disease virus

Kyasanur forest virus disease (KFD) is a major public health concern in India. Its etiology KFD virus causes haemorrhagic fever with severe sequelae in humans. Due to continuous spatiotemporal expansion of KFD in last decade, the incidences of positive cases have been increasing in both humans and primates. Early diagnosis is of prime importance for disease management and epidemiological containment. In the present study, the highly immunogenic Envelope Domain III (EDIII) antigen was produced using prokaryotic expression system with an yield of 8 mg/L. The protein was purified using affinity chromatography and confirmed for its immuno-reactivity by western blot and UPLCMS/MS analysis. The recombinant EDIII was used as an antigen for the standardization of ELISA to detect anti KFD IgM antibodies in humans. The ROC curve was prepared to set the optimum cut-off OD for the assay. The comparative evaluation of the assay with a reference MAC ELISA revealed 86.96% concordance, 82.22% sensitivity and 91.48% specificity. Inter-rater agreement was performed with kappa index revealing significant agreement between the assays. This is the first study using safe recombinant protein antigen-based detection of anti KFDV antibodies in humans. This simple and scalable ELISA assay will be applicable for large scale screening of samples for combating the emerging threats of KFD in newer territories.

Laboratory Parameters of Dengue Infection in a Medical College Hospital

Introduction &amp; objective: Dengue is an acute illness caused by Aedes mosquito, commonly caused by Aedes aegypti. It is an endemic disease in South East Asian countries especially in Bangladesh. In 2019 a outbreak occurred in Bangladesh. The objective of the study was to see the laboratory parameter and outcome of dengue fever in a tertiary medical college hospital. Early diagnosis and laboratory investigations is essential to prevent the mortality associated with this disease. Materials &amp; Methods: This prospective study was conducted on dengue ward at Khulna medical college hospital from 1 July 2019 to 31 December 2019. The diagnosis of dengue infection was confirmed by serology. Sera were processed by dengue ELISA. Investigations like haemoglobin estimation, haematocrit, platelet count, total count; differential leukocyte count, peripheral smear, coagulation profile were performed. Results: Total number of cases was 98. Of them 62 (63.2%) was male and 36 (36.73%) was female. Fever was most common (100%) manifestation and duration of fever ranges from 5 to 8 days with a mean duration of 6.31 (± 0.95) days. Blood for NS1 was positive in 90 (91.83% ) cases; most cases 71 (72.44%) become positive between 2nd to 4th day. 90 (91.83%) cases developed thrombocytopenia and it starts at 5th day in 45 (45.91%) cases. Anti IgM was positive in 4 (4.08%) cases; 2 at 4th day and 2 at 5th day of illness. IgG was positive in case at 8th day of illness. Conclusion: These findings help physicians in early diagnosis of dengue by suspecting these features as of dengue and can prevent morbidity and mortality associated with dengue. Medicine Today 2022 Vol.34(1): 61-64

The association of human cytomegalovirus with biomarkers of inflammation and immune activation in breast cancer.

The presence of human cytomegalovirus (HCMV) in breast cancer has been reported, suggesting a potential association between HCMV infection and breast carcinogenesis. To evaluate the association between HCMV infection and immune activation and inflammatory markers in breast cancer. HCMV DNA was detected from all patients using real-time PCR, Anti HCMV IgM and IgG antibodies were measured. IL-17 and IL-22 concentrations were detected by ELISA. Assessment of NLR and PLR was done, and cell proliferation was assessed using MTT assay. The results revealed a significantly increased prevalence of anti-HCMV IgG and HCMV DNA in patients compared to both benign and control groups where positive HCMV prevalence was significantly associated with vascular invasion, proliferation rate, high neutrophil-to-lymphocyte ratio (NLR), and elevated IL-17 serum level. Furthermore, we demonstrated that increased serum IL-17 in patients was markedly associated with tumor stage, vascular invasion, and high NLR. It can be concluded that HCMV infection may have vital roles in breast cancer pathogenesis. Moreover, altered peripheral blood cells and cytokines may result in disordered immune response in breast cancer patients.

An Investigation Profile in Pediatric Patients Suspected of Acute Hepatitis A

Hepatitis A virus (HAV) infection is one of the important causes of hepatitis in developing countries. It is transmitted through feco-oral route. This poses major health problems in children. Serum samples of the patients from pediatrics department suspected of acute viral hepatitis presenting with clinical features like fever, nausea, vomiting, and jaundice were collected and analyzed by using commercially available mini VIDAS 307 (Biomeriux) to detect anti HAV IgM antibodies. A nine months prospective study was conducted in the Department of Microbiology and immunology of Shri Guru Ram Rai Institute of Medical and health science Dehradun, India. Of the 47 serum samples received during the study period, 23 (48.93%) were found to be positive for anti-HAV IgM (VIDAS 307 Biomeriux). The prevalence was found to be more in male patients than in female patients, that is,14 (60.8%) and 9 (39.13%) respectively, and was predominantly seen in the 5–10 years of the age group (60%). Peak cases were found in the month of October and November. The study showed a high positivity (48.93%) of HAV in this sub-Himalayan region indicating the role of preventive measures including safe drinking water, improved sanitation and vaccination in preventing infection. Thus, detection of anti-HAV IgM becomes the cornerstone entity for diagnosis.

English

BACKGROUND Hepatitis A &amp; E viruses are enterically transmitted viruses responsible for acute viral hepatitis in developing countries. Hepatitis A virus (HAV) has worldwide distribution while hepatitis E virus (HEV) is restricted to tropical countries. HAV affects infants and young children while HEV affects older children and young adults. This study was conducted to determine the seroprevalence of HAV and HEV and their co-infection in patients presenting with acute viral hepatitis (AVH). METHODS 200 sera of patients suffering from suspected acute viral hepatitis (AVH) attending tertiary care rural hospital JNMC Sawangi, Wardha were included in the study. Serum samples were tested for IgM anti HAV and IgM anti - HEV for the detection of acute hepatitis A and acute hepatitis E using commercially available ELISA kit &amp; Immunochromatography test (ICT) for comparison of ELISA and ICT. RESULTS The overall seroprevalence of HAV and HEV infection was found to be 19 %. The seroprevalence of HAV infection was found to be 13 %, HEV infection 5 % and HAV - HEV co - infection 1 %. HAV infection in males and females was found to be 16.07 % and 9.09 % respectively &amp; HEV infection was found to be 5.35 % and 4.54 % respectively. In case of detection of HAV IgM, two serum samples were negative by ICT but positive by ELISA &amp; for HEV IgM, one serum sample was negative by ICT but positive by ELISA. The sensitivity, specificity, positive predictive value and negative predictive value of HAV IgM was found to be 96 %, 98.8 %, 92.30 % &amp; 99.43 % and for HEV IgM was found to be 90 %, 99.47 %, 90 % &amp; 99.47 % respectively. CONCLUSIONS It is evident from this study that infection with enteric hepatitis viruses is not infrequent. Findings from this study emphasize the need to establish regular seroepidemiological surveys to keep track of epidemiology of these viruses. Results from rapid tests are comparable to ELISA with additional advantage of ease of interpretation and neither time consuming nor requiring special instruments. KEY WORDS Hepatitis A, Hepatitis E, Seroprevalence

ASSOCIATION OF SEROPOSITIVITY FOR CMV IGM AND IGG WITH ADVERSE PREGNANCY OUTCOME

Objective: To find out the prevalence of antibodies against cytomegalovirus and their association with adverse pregnancy outcome in women, at Isra University Hospital Hyderabad.&#x0D; Study Design: Cross-sectional study.&#x0D; Place and Duration of Study: Department of Gynaecology &amp; Obstetrics, Isra University Hospital (IUH) Hyderabad, from Jan to Jun 2018.&#x0D; Methodology: Cytomegalovirus IgM and IgG antibodies in 305 women of reproductive age group were measured using the Enzyme Linked Fluorescent Assaysystem kit.&#x0D; Results: The combined positivity of anti cytomegalovirus IgG and IgM antibodies was 93 (30.40%). About 37 (37.37%) and 28 (34.14%) women were found to be seropositive for cytomegalovirus antibodies who had history of spontaneous miscarriage and recurrent miscarriages respectively.&#x0D; Conclusion: Higher association of seropositivity for cytomegalovirus IgM and IgG with bad obstetrical history and low economy was found.

Hepatitis E viral infection in pregnant ladies, a challenge for obstetricians and physicians

Background: Hepatitis E Virus (HEV) is the major cause of morbidity and mortality in pregnancy. The virus was detected about three decades ago. The incidence and severity during pregnancy vary widely around the world.Material &amp; Methods: This observational retrospective study was conducted from March 2019 to February 2020 in Obstetrics &amp; Gynecology Department of Lady Aitchison Hospital, Lahore. 70 pregnant women with hepatitis E were subjected to detailed history taking. The diagnosis was based on positive Hepatitis E IgM (Anti HEV IgM) antibody on laboratory test in current pregnancy in antenatal period up to 42 days post partum were included.Results: Out of 70 pregnant women with Hep E IgM+ve, 3 (4.3%) women had Obstetrical hysterectomy as complications, with mean age of 26.0 ± 4.69 years. The mean gestational age when infection occurred was 31.12 ± 5.18 weeks. Among all females 9 (13%) patients expired due to fulminant hepatic failure, 12 (17.14%) females gave birth via spontaneous vaginal delivery (SVD),17(24.2%) females gave birth via C-Section. 30 (42.8%) females were manage conservatively.Conclusion: Hepatitis E viral infection in pregnant females is a life threatening condition. The study showed that pregnant women with jaundice and hepatitis E virus infection had a high mortality rate during third trimester. Early diagnosis and management will help in the control of the disease and prevents its complications.  Â

Severity of dengue illness and presence of anti DV IgG in serum of laboratory confirmed dengue cases.

Presence of dengue is reported from India since 1960s. Secondary dengue infection may be more severe than primary, hence, distinction between primary and secondary dengue is essential. A way to detect secondary dengue is demonstration of anti DV IgG in patients' serum. In this study we explored the association of dengue severity with anti DV IgG positivity. Laboratory confirmed cases of dengue (positive for anti DV IgM/ NS-1 Antigen/ DV -RNA), presenting to the hospital within 7 days of illness, were consecutively enrolled for a period of one month (September 1-30, 2018) and were tested for anti DV IgG in their serum. All PCR positive samples were serotyped. Cases positive for anti-dengue IgG were labeled as secondary cases. Clinical details were collected to assess the severity of illness. Association of dengue severity with anti DV IgG positivity was calculated. Of the 128 dengue positive cases, 89 (69.5%) were anti DV IgM positive, 72 (56.3%) were Dengue NS-1 positives and 37 (28.9%) were DV-RNA positive. Only 39 (30.5%) cases were having detectable anti-dengue IgG in their serum (secondary dengue). Anti-dengue IgM positivity was significantly higher in secondary dengue cases. No association of anti DV IgG positivity was seen with severity of dengue illness. No association of IgG positivity with severity of illness was seen. D4 serotype is first time reported from Uttar Pradesh, India.

Prevalence of HAV and HEV in Patients Presenting with Acute Viral Hepatitis in Silchar Medical College & Hospital

Background: Hepatitis A virus (HAV) and Hepatitis E virus (HEV) are both enterically transmitted, resulting in acute viral hepatitis (AVH) in developing countries. Both are self-limiting viral infection, transmitted by feco-oral route. Exposure rate of HAV and HEV over a period are different in different parts of country. Aim: This study was done to determine prevalence of HAV and HEV in patients presenting with AVH and the co-infection of HAV and HEV in these patients. Materials and Methods: A cross sectional study of 2 years duration was conducted in the Department of Microbiology, SMCH, Silchar. A non random sampling of 200 patients presenting with AVH was considered in the study. On the basis of history, serum sample were analyzed for IgM anti HAV and IgM anti HEV for the detection of HAV and HEV, respectively using commercially available ELISA kits. Results: The prevalence of HAV and HEV co-infection in patients with acute viral hepatitis was 32.5% (65 cases). HAV and HEV were most prevalent between 20 to 35years age group. Hepatitis A samples only 60 were positive and Hepatitis E samples only 5 were positive. Conclusion: Almost all HAV and HEV infectious are spread by feco-oral route. The best means of reducing the spread of infection is by promoting simple measures of personal and community hygiene such as hand washing before eating and after defecation, the sanitary disposal of excreta which will prevent contamination of water, food and milk, during epidemics, boiled water should be advocated for drinking purposes.

Improved Early Detection of Focal Brucellosis Complications with Anti-Brucella IgG.

To evaluate the associations of inflammatory factors and serological test results with complicated brucellosis, we recruited 285 patients with a diagnosis of brucellosis between May 2016 and September 2019. The patients were subsequently classified into two groups according to the presence of complications. We collected demographic and clinical information and routine laboratory test results in addition to anti-Brucella IgG and IgM levels. Anti-Brucella IgG and IgM were uniformly tested using enzyme-linked immunosorbent assays (ELISAs) in this study. Among the 285 patients with brucellosis, 111 (38.95%) had complicated brucellosis. Osteoarthritis occurred more often in the subacute and chronic stages than in the acute stage (P = 0.002). Genital infection occurred more frequently in the acute stage than in the other stages (P = 0.023). Fever was not frequently observed in complicated cases (P < 0.001). The erythrocyte sedimentation rate (ESR) and the C-reactive protein (CRP) and anti-Brucella IgM and IgG levels were higher in complicated-brucellosis patients than in uncomplicated-brucellosis patients (P < 0.001). Anti-Brucella IgG, with an area under the curve of 0.885 (95% confidence interval [CI], 0.847 to 0.924), was the most robust indicator of complicated brucellosis. Positive culture, anti-Brucella IgM, the ESR, and CRP could be considered indicators, but their efficacy was weaker than that of IgG. In conclusion, a high ESR, high CRP, high anti-Brucella IgM and IgG levels, and positive culture were indicators of complicated brucellosis; among these, anti-Brucella IgG was the most robust biomarker.

Role of Congenital Cytomegalovirus Infection and Protein 53 in Neural tube Defect

Background: Neural tube defects (NTDs) are the greatest shared severely disabling birth defects, they resulted from compound and many etiologies in which both hereditary, life style and ecological factors appear to be convoluted. Objective: To study the role congenital cytomegalovirus and the biomarker Protein 53 (p 53) in pathogenesis of neural tube defects. Patients and Methods: This is a cross sectional comparative study done on 47 mothers of different age group and their babies who have NTD and 47 mothers with their healthy babies at AL-Batool Teaching Hospital/ Diyala/ Iraq from Sept 2017 to May 2018. Rapid serological test to detect anti- CMV IgG and IgM Abs for all case and control groups and their mothers, together with ELISA test to detect P53. The statistical analysis of data was achieved by using SPSS V.20. Results: There was high rate of anti- CMV IgG Abs in NTD cases and their mothers ( n= 10,21.3%, p value= 0.08), out of them, one case had positive anti-IgM Abs together with his mother, whereas in control group, anti CMV IgG was positive in 4(8.5%)in both mothers and their healthy babies with no anti IgM detected. P53 titer is higher to a significant level ( p<0.05) in the mother and their NTD affected baby than control groups ( corresponding babies and mothers). Conclusion: CMV virus might be an important cause of NTD and p53 is a likely to be involved in pathogenesis of NTD.

Antibodies to Zika virus, Campylobacter jejuni, gangliosides in Guillain Barre syndrome. A prospective single centre study from southern India

The objective was to study the presence of antibodies against Zika virus, Camplyobacter jejuni and gangliosides in patients with Guillain Barre syndrome. Ninety consecutive patients (age more than 12 years) with GBS admitted to a tertiary care centre in southern India were included in this study. Data on the clinical manifestations, Nerve conduction studies and response to therapy were collected. The following tests were done in stored serum samples - Anti-Zika virus (IgM) antibodies, Anti-Campylobacter jejuni (IgG) antibodies and Anti-ganglioside antibodies (IgG). Those samples which were positive to Anti Zika antibodies were tested for conventional PCR for Zika virus, IgM antibodies against Dengue and Japanese encephalitis. Of the 90 patients, 3 died and 8 had persistent weakness. AIDP was the most common type of GBS (56.7%). Anti-ganglioside antibodies were present in 62.2% with GT1b being most common. Anti-C.jejuni antibodies were present in 46.6%. Anti-Zika antibodies (IgM) were present in 14 patients (15.5%). Four of these patients also had anti-Dengue antibody (IgM) positivity. This is one of the largest study on GBS from India and the first one to report on the presence of Zika antibodies form this geographical area. Our study had high prevalence of anti C.jejuni and anti ganglioside antibodies. Evidence of recent ZIKV infection as evidenced by anti IgM antibodies was present in 14 patients with 4 of them tested positive for anti-Dengue IgM antibody. Whether this represents cross reaction with Dengue or prior/co- infection with Dengue could not be addressed in this study.

Abstract 876: CD40-induced growth inhibition of Burkitt lymphoma: A possible role for NADPH oxidase by upregulation of p67phox

Abstract CD40 stimulation together with IL4 has been shown to promote growth of normal B cells in vitro. In contrast to this, CD40 stimulation in several B cell lymphoma cell lines has been demonstrated to be growth inhibitory. Though the mechanism is not completely known, CD40-stimulated cells exhibit increased levels of reactive oxygen species (ROS). In B lymphocytes, a major source of ROS production is the NADPH oxidase (NOX) that is comprised of a membrane-bound Nox2 catalytic subunit and four cytosolic regulatory components, with the p67phox cytosolic protein being the activator factor. Upon stimulation, by either cross linking of the B-cell receptor or following treatment with PMA, the active NADPH oxidase generates superoxide. We have investigated the role of NADPH oxidase in CD40-mediated growth inhibition in Burkitt lymphoma (BL) cells. For this study, we first evaluated the effect of CD40 stimulation on the expression of Nox2 and p67phox in BL cells that demonstrate different levels of endogenous Nox2 and p67phox. Accordingly, Daudi and Raji cell lines, with endogenously high Nox2 and low p67phox, NAMALWA cells which have high Nox2 and no p67phox expression, and Ramos cells that contain no Nox2 or p67phox were analyzed. Treatment with anti-CD40 or CD40 ligand (CD40L) for 24 h induced up-regulation of p67phox in Daudi, Ramos, and NAMALWA cells, but not in the Raji line. No significant change in the expression of Nox2 was observed. Additionally, Daudi, Ramos and NAMALWA cells showed higher production of ROS after CD40 stimulation and activation with anti-IgM or PMA, in a chemiluminescence assay. To determine whether the change in ROS level was related to the upregulation of p67phox, Daudi cells that express either stable p67phox shRNA or control shRNA were generated. Treatment with CD40L for 24 h demonstrated significant ROS production only in control shRNA and not in the p67shRNA cells, confirming that the ROS production is NOX2-p67phox mediated. This enhanced ROS production was observed following treatment with CD40L alone and was enhanced further in conjunction with anti-IgM or PMA. However, cells with the p67phox shRNA showed detectable level of ROS only after activation with anti IgM, albeit to much lower levels than the control. Co-treatment of Daudi cells with anti-CD40 or CD40L and IL4 further increased the expression of p67phox. Although the basal level of ROS in the cells was unchanged upon co-treatment, the ROS generated following stimulation with anti-IgM or PMA was significantly enhanced. Preliminary MTT studies confirmed the growth inhibition of CD40-stimulated Daudi cells but not of Raji cells, suggesting a role for p67phox-dependent, ROS-related inhibition of lymphoma cell proliferation. Experiments are currently ongoing to delineate further the role of CD40 stimulation and Nox2-p67phox mediated ROS in affecting the growth of Burkitt lymphomas and signaling pathways downstream of CD40. Citation Format: Mazal Iris Dahan, Smitha Antony, Agnes Juhasz, Guojian Jiang, Mariam Konate, Jiamo Lu, Jennifer Meitzler, YongZhong Wu, Krishnendu Roy, James H. Doroshow. CD40-induced growth inhibition of Burkitt lymphoma: A possible role for NADPH oxidase by upregulation of p67phox [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 876.