Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device.

LAMP diagnosis of malaria is simple and cost-effective with acceptable sensitivity and specificity as compared to standard diagnostic modules such as microscopy, RDTs and nested PCR, and thus its deployment for onsite screening of malaria in resource-limited regions is under consideration. However, the requirement of an electricity-operated dry bath and bulky read-out unit is still a major concern. In an effort to simplify this limitation, we have developed a portable LAMP device and fluorescence readout unit which can be used in the rapid point-of-care diagnosis of malaria. We have developed a point-of-care diagnostic LAMP device that is easy to operate by a mobile application, and the results can be quantified with a fluorescent readout unit. The diagnostic performance of the device was evaluated in 90 P. falciparum-infected clinical isolates stored at 4°C for 6-7 years and 10 freshly collected isolates from healthy volunteers. The LOD and quantitative ability of LAMP in estimating parasitemia levels were revealed with laboratory-grown P. falciparum strain (3D7). The LAMP assay performed in our device was exclusive for P. falciparum detection with sensitivity and specificity determined to be 98.89% and 100%, respectively, in clinical isolates. The LOD was documented to be 1 parasite/µl at the cut-off ADC value of 20. Parasite density estimated from ADC values showed concordance with microscopically determined parasite density of the cultured P. falciparum 3D7 strain. The LAMP assay performed in our device provides a possible portable platform for its deployment in the point-of-care diagnosis of malaria. Further validation of the quantitative ability of the assay with freshly collected or properly stored clinical samples of known parasitemia is necessary for field applicability.

A Study on the Temporal Trends in the Etiology of Cirrhosis of Liver in Coastal Eastern Odisha.

BACKGROUND: The etiology of cirrhosis of liver is known to change with time due to various factors including awareness, preventive interventions, and lifestyle changes in society. However, there is scarce Indian data available about temporal trends in etiology of cirrhosis of liver. Hence, the aim of this study was to study the temporal trends in the etiology of cirrhosis of liver. MATERIALS AND METHODS: This is a retrospective study conducted in the Department of Gastroenterology, Srirama Chandra Bhanja Medical College and Hospital, Cuttack, from January 2005 to December 2017. Data were collected from hospital records of all patients admitted to the Gastroenterology unit. A Poisson regression model was used to compare the hospitalization rate for different etiologies of cirrhosis of liver. All data were analyzed using Stata version 5.1 software. RESULTS: A total of 4,331 hospitalized patients of cirrhosis of liver were included in the analysis, of whom 2,742 (63.3%) had alcohol-related cirrhosis, 858 (19.8%) had viral hepatitis-related cirrhosis, and 731 (16.9%) had cirrhosis of liver due to nonalcohol and nonviral causes. The proportion of alcohol-related cirrhosis was increased by 26% from 2005 to 2017 (RR 1.26, p for trend <0.001). Though there were minimal ups and downs observed in the admission rate of viral hepatitis-related liver cirrhosis during later years, this was remarkably reduced by 73% (RR 0.27, p for trend <0.001) in the year 2017 at the end of the study. Similarly, the proportion of cirrhosis due to nonalcohol and nonviral causes decreased by 26% (RR 0.74, p for trend <0.001) by 2017. CONCLUSION: Alcohol is the most common cause of cirrhosis of liver and the burden of alcohol-related cirrhosis is significantly increasing in comparison to other causes including viral infection, nonalcoholic steatohepatitis (NASH), and autoimmune hepatitis. HOW TO CITE THIS ARTICLE: Mishra D, Dash KR, Khatua C, et al. A Study on the Temporal Trends in the Etiology of Cirrhosis of Liver in Coastal Eastern Odisha. Euroasian J Hepato-Gastroenterol 2020;10(1):1-6.

Validation of International Club of Ascites subclassification of stage 1 acute kidney injury in chronic liver disease.

BACKGROUND AND AIM: Acute kidney injury (AKI) commonly occurs in patients with chronic liver disease (CLD). As per the International Club of Ascites, AKI is classified into three stages; stage 1 has recently been divided into subgroups 1A and 1B. We performed a prospective study to validate the association between subgrouping and outcome. METHODS: This study was conducted using decompensated cirrhosis (DC) patients hospitalized in the Gastroenterology ward between August 2016 and May 2018. Demographic, clinical, and laboratory parameters were compared between AKI 1A and AKI 1B patients. The duration of hospitalization and outcome were compared. RESULTS: A total of 528 subjects were enrolled; 296 (56.1%) had AKI, and of them, 61.48% (n = 182) had stage 1, 20.95% (n = 62) had stage 2, and 17.57% (n = 52) had stage 3 AKI. Of the enrolled patients, 100 (54.94%) had early (AKI 1A) and 82 (45.06%) had late stage 1 AKI (AKI 1B). Patients with AKI 1B had higher total leucocyte count, total bilirubin, serum urea, serum creatinine (SCr), model for end-stage liver disease (MELD), MELD-Na+, and child-turcotte-pugh (CTP) score and decreased serum albumin than AKI 1A. The prevalence of hepatorenal syndrome (HRS), acute on chronic liver failure (ACLF) were higher in AKI 1B patients, and they had a prolonged hospital stay compared to AKI 1A patients. Furthermore, AKI 1B patients had significantly lower survival both at 28 days and 90 days. CONCLUSION: Our study validates the subclassification of stage 1 AKI. Patients with AKI 1B more often progress to higher AKI stages with significantly lower 28-day and 90-day survival rates. Results justify subclassification and suggest the need for early intervention. The small increase in SCr should be viewed with caution in AKI stage 1A.

A prospective study on incidence, risk factors, and validation of a risk score for post-infection irritable bowel syndrome in coastal eastern India.

BACKGROUND AND AIM: Post-infection irritable bowel syndrome (PI-IBS) can occur following acute gastroenteritis (AGE). This study was designed to evaluate the incidence and risk factors of PI-IBS following AGE and to validate a PI-IBS risk score. METHODS: This prospective study was performed between September 2014 and October 2016 on AGE patients by documenting their AGE severity and following up after 3 and 6 months to study the development of IBS (ROME III criteria). The risk score was calculated for all the subjects, and its discrimination ability was tested. RESULTS: Out of 136 hospitalized AGE patients, 35 developed PI-IBS after 6 months. The factors associated with PI-IBS were younger age, longer duration of AGE, anxiety, depression, abdominal pain, bloody stool, vomiting, fever, family history of IBS, and positive stool culture (univariate analysis); however, on multivariate analysis, younger age (adjusted odds ratio [AOR] 0.5; p 0.03), prolonged duration of AGE (AOR 8.6; p 0.01), and abdominal cramps (AOR 2.1; p 0.02) were the independent factors influencing its occurrence. PI-IBS occurred even after infection with Vibrio cholerae. The PI-IBS risk score was significantly higher in patients who developed PI-IBS (72.4 ± 14.48 vs. 31.56 ± 20.4, p-value < 0.001); score > 50 had a sensitivity and specificity of 91.4% and 84.2%, respectively. CONCLUSION: One fourth of AGE patients developed PI-IBS after 6 months. Factors influencing its development were younger age, long duration of AGE, and abdominal pain. The PI-IBS risk score had good predictive accuracy in our population.

Effect of Plasmodium falciparum infection on blood parameters and their association with clinical severity in adults of Odisha, India.

The extent of abnormalities in blood indices and their subsequent effects on clinical severity in malaria differ among populations of different endemicity. However, these alterations have not been well investigated in Odisha, India and their prognostic implications in the context of multi-organ dysfunction (MODS) in severe malaria (SM) are not identified so far. The present study was carried out in 200 adult patients each from uncomplicated malaria and severe malaria groups to examine whether host haematological and biochemical parameters in Plasmodium falciparum infection can act as diagnostic marker for SM in adults patients of Odisha. The results showed thrombocytopenia as a potential risk factor for SM irrespective of disease features with least median platelet counts observed in patients with MODS (Platelet count: 144.5, P = < 0.0001) compared to mild malaria. Logistic regression analysis identified anemia (<10 mg/dl) as independent predictor of MODS (OR = 12.78, 95% CI = 4.93-33.2). The prognostic utility of thrombocytopenia (platelet count: ≤100,000/µl) as marker of MODS was largely modulated by hemoglobin and blood glucose level. Co-existence of hypoglycemia and thrombocytopenia was also observed. Our study revealed changes in blood indices such as low platelet, hemoglobin and blood glucose during falciparum infection in adults can be used as diagnostic criteria for predicting SM in combinations. The study also provides important clue for plausible hypoglycemia mediated platelet necrosis and clearance. Further studies in different endemic regions need to be conducted for validation of these findings and their implication as criteria for diagnosing SM in adults.

Acute Kidney Injury at Admission Is a Better Predictor of Mortality than Its Persistence at 48 h in Patients with Acute-on-chronic Liver Failure.

Background and Aims: Acute kidney injury (AKI) occurs commonly in patients with acute-on-chronic liver failure (ACLF). However, there are scant data regarding the impact of AKI on survival in ACLF. We performed a prospective study to evaluate the impact of AKI on survival in ACLF. Methods: This study was conducted in ACLF patients hospitalized in the Gastroenterology Department of Sriram Chandra Bhanja Medical College (India) between October 2016 and February 2018. Demographic, clinical and laboratory parameters were recorded, and outcomes were compared between patients with and without AKI and between patients with persistent AKI and AKI reversal at 48 h. Results: We screened 439 chronic liver disease patients as per the Asian Pacific Association for the Study of the Liver criteria and found that 113 (25.7%) of them had ACLF and 78 (69%) of them had AKI as per the Acute Kidney Injury Network criteria. ACLF patients with AKI had reduced 28-day survival (44.9% vs. 74.3%; p = 0.004) and 90-day survival (25.6% vs. 51.4%; p = 0.007), in comparison to ACLF patients without AKI. However, when comparison was made between AKI reverters and AKI persisters at 48 h, survival was comparable for both at 28 days and 90 days. Further, about one-tenth of ACLF patients with AKI died within 48 h of hospitalization. Conclusions: Over two-thirds of ACLF patients had AKI. Although ACLF itself is a predictor of reduced survival, a very small increase in serum creatinine further worsens survival. Importantly, AKI at admission is a better predictor of early mortality in ACLF patients since recovery from AKI at 48 h does not improve survival.

Genetic predisposition of variants in TLR2 and its co-receptors to severe malaria in Odisha, India.

Although the role of TLRs signalling in malaria pathogenesis is well established, contribution of individual TLR to clinical outcome of malaria still remains inconclusive. Given the importance of TLR2 and its co-receptors in recognising distinct structural forms of key malaria toxins and mediating innate immune response, it is essential to delineate their genetic contribution. Variants in TLR1 (I602S) and TLR6 (P249S) were genotyped by PCR-RFLP methods, and TLR2 (I/D) was genotyped by PCR in 200 samples each from uncomplicated malaria (UM) and severe malaria (SM). Further, SM was categorised into its sub-clinical groups (CM and NCSM or SOD and MODS) and analysed. The results showed the PP genotype of TLR6 (P249S) to be significantly more common in UM (P < 0.0001), whereas the 'SS' genotype was the risk factor for SM including its sub-clinical categories. The TLR1 (602S) and TLR2 (D) variants were significantly high in patients with CM; however, negative LD was observed between TLR2 and TLR6 in NCSM and MODS. Haplotype analysis showed significantly high frequency of I-I-S haplotype in all forms of subclinical SM and was associated with low parasite load in SM (P = 0.013). The haplotypes I-D-S and S-I-P were significantly high in SOD and CM, respectively. The TLR6 '249S' variant appeared to be the dominant determinant for genetic predisposition to SM and that its association with either TLR2 'D' or TLR1 '602S' modulates for CM development. The present study opens up several new avenues for their exploration and validation in future studies in different global settings for malaria.

Influence of common variants of TLR4 and TLR9 on clinical outcomes of Plasmodium falciparum malaria in Odisha, India.

BACKGROUND: In malaria, the toll-like receptors (TLRs) have recently emerged as major player of innate immunity. However, implication of TLR variants on clinical manifestations of malaria is conflicting. The present study aims to provide relevant information of growing interest in understanding the role of TLR4D299G, TLR9T-1237C and TLR9T-1486C polymorphisms on clinical outcomes of malaria. METHODS: We genotyped TLR4D299G, TLR9T-1237C and TLR9T-1486C polymorphisms by PCR-RFLP methods and subsequently analyzed in 200 uncomplicated patients and 200 severe patients. Further, the severe malaria categorized into sub-clinical groups such as cerebral malaria (CM), non-cerebral severe malaria (NCSM), single organ dysfunction (SOD) and multi-organ dysfunctions (MODS) are analyzed. RESULT: The TLR9-1237CC genotype was observed at significantly low frequency in MODS (p=0.0008), while in heterozygous state (TC) it was proportionately more frequent in SOD (p=0.087) as compared to mild malaria. The TLR9T-1486C heterozygote was more common in all categories of severe malaria. However, pair wise LD analysis revealed significant linkage between T-1237C and T-1486C, whereas haplotype analysis showed significantly low frequency of C-T haplotype in CM (p=0.005, pc=0.02) and high frequency of T-C haplotype in NCSM as compared to mild malaria. CONCLUSION: Although TLR9-1237C could be a risk factor for severe malaria in heterozygous state, negative association of CC genotype with MODS warrants caution of segregating severe malaria into its sub-clinical groups while interpreting data. Further, clinical outcome in malaria was observed to be apparently modulated by LD between TLR9 promoter variants.