Laboratory

Urinary N-acetyl-beta-d-glucosaminidase levels in diabetic adults Eiya Bibiana Omozee, Enajit Ibiene Okaka, Andrew Efosa Edo, Leonard Fedinard Obika Journal of Laboratory Physicians 2019 11(1):1-4 Diabetes mellitus (DM) is known to be one of the most common causes of end-stage renal disease. The disease is usually not detected on time, because of the large functioning reserve of the kidney. Currently used markers (serum creatinine, creatinine clearance, urea, and electrolytes) remain relatively normal even when more than 50% of the renal nephron is not functioning. The aim of this study was to determine the level of urinary N-acetyl-beta-d-glucosaminidase (NAG) in diabetic adults in comparison with some currently used markers. A total of 56 diabetic patients between the ages of 23 and 63 were used for this study and 30 nondiabetic between the ages of 18 and 62 were used as control. The diabetic patients were classified into three groups based on how long they have been diagnosed: <2 years (25), 2–5 years (30), and >5 years (25). Spot midstream urine samples were collected into sterile containers, and blood samples were collected into plain tubes. All the analyses were done spectrophotometrically. Creatinine clearance was calculated using the Cockcroft–Gault Equation. There was a significant increase (P < 0.01) in NAG values of 2–5 years and above 5 years and control. The urinary microalbumin concentration of controls was significantly different (P < 0.05) only with those who have had DM for <2 years. Urinary creatinine concentration of control was significantly higher (P < 0.05) than values of all the diabetic groups. There was a significant increase (P < 0.01) in creatinine clearance of control group and those who have had DM for <2 years. It is thus concluded that urinary NAG can be used as an early marker in the diagnosis of diabetic nephropathy since urinary NAG increases first before the other markers analyzed in this current study begins to increase.

Clinical profile and determinants of short-term outcome of acute kidney injury: A hospital-based prospective study from Northeastern India Prasanta Kumar Bhattacharya, Akash Roy, Md. Jamil, Bhupen Barman, Subrahmanya V Murti, Patrick R Marak Journal of Laboratory Physicians 2019 11(1):5-10 CONTEXT: Acute kidney injury (AKI) is an outcome of multiple etiologies and is mostly reversible. Data on its incidence and outcome, particularly from India, are limited. AIMS: To study the etiology, clinical profile, and short-term prognosis in AKI. SETTINGS AND DESIGN: A hospital-based prospective observational study on AKI. SUBJECTS AND METHODS: Seventy-five AKI patients diagnosed by Acute Kidney Injury network criteria were selected. Patients with preexisting chronic kidney disease were excluded. STATISTICAL ANALYSIS USED: Data were compiled using the Statistical Package for the Social Sciences version 17. Regression analysis was done for determining the association of various variables for mortality. P < 0.05 was considered statistically significant. RESULTS: The mean age of patients was 41.09 ± 16.17 years with a male:female ratio of 1.42:1. Comorbidities were present in 37.3%, with diabetes mellitus (10.6%) and chronic liver disease (10.6%) being the most common. Fever was the most common (40%) presenting symptom, followed by oliguria (25.8%). Infection was the most common cause of AKI (56%), with sepsis in 26.7% followed by acute gastroenteritis in 17.3%. Pneumonia was the primary focus in 50% of cases with sepsis. Mean serum creatinine and urea at admission were 2.37 ± 0.90 and 92.44 ± 39.67 mg/dl, respectively. Serum creatinine rose progressively to 2.96 ± 1.18 and 3.26 ± 1.56 mg/dl at 24 and 48 h, respectively, since hospitalization. Majority of the cases (73.3%) were nonoliguric. Hemodialysis was necessary in 24% of cases. Mean hospital stay was 8.16 days. In-hospital mortality was 24%. Among survivors, 92.9% had complete renal recovery on discharge. Sepsis, need for hemodialysis, urea >100 mg/dl, and peak serum creatinine >3 mg/dl were contributors to mortality (P < 0.01). CONCLUSIONS: Infections, especially sepsis, were the most common cause of AKI. Hemodialysis was required in one-quarter of the patients. Sepsis, need for hemodialysis, and high creatinine were associated with a significantly higher mortality.

Serological evidence of human leptospirosis in patients with acute undifferentiated febrile illness from Uttarakhand, India: A pilot study Mohit Bhatia, Pradeep Kumar, Pratima Gupta, Puneet Kumar Gupta, Minakshi Dhar, Deepjyoti Kalita Journal of Laboratory Physicians 2019 11(1):11-16 CONTEXT: To the best of our knowledge, there are no reports of serological evidence of human leptospirosis from Uttarakhand state in India. AIMS: The aim of this study was to screen for serological evidence of leptospirosis in patients with acute undifferentiated febrile illness at a tertiary care teaching hospital in Uttarakhand. SETTINGS AND DESIGN: A pilot study was conducted from March to November 2017. SUBJECTS AND METHODS: Fifty-three adult patients who presented in Medicine outpatient Department with a history of fever of ≥7 up to 14 days duration with or without other associated symptoms such as a headache, rashes, myalgia, arthralgia, and conjunctival suffusion were enrolled in the study using convenience sampling technique. Blood samples of these patients were collected and subjected to peripheral smear examination for malaria parasites, dengue immunoglobulin M (IgM) immunochromatographic card test, IgM Typhidot, Leptospira and Scrub typhus IgM ELISA, respectively. Aerobic blood culture was performed in 24 cases. Relevant clinico-epidemiological details were obtained as per the pro forma formulated in accordance with the modified Faine's criteria. STATISTICAL ANALYSIS USED: Descriptive statistics. RESULTS: The study population consisted of 50.94% of males and 49.06% of females with a mean age ± standard deviation of 34.2 ± 15.2 years. Fifty febrile patients had additional symptoms of which myalgia was the most common (81.1%) followed by arthralgia (22.6%). Peripheral smears of all patients were negative for malaria parasites. Dengue and Typhidot IgM positivity was observed in two and eight patients, respectively. Six and five patients were tested positive by leptospira and scrub typhus IgM ELISA, respectively. Salmonella Typhi was isolated from blood sample of only one patient. Serum samples of two patients showed dual positivity. All six leptospira seropositive patients satisfied modified Faine's criteria. CONCLUSIONS: Leptospirosis is a seemingly unexplored infection in Uttarakhand and should be considered as a differential diagnosis in patients with acute undifferentiated febrile illness.

Detection of biofilm among uropathogenic Escherichia coli and its correlation with antibiotic resistance pattern Rashmi M Karigoudar, Mahesh H Karigoudar, Sanjay M Wavare, Smita S Mangalgi Journal of Laboratory Physicians 2019 11(1):17-22 BACKGROUND: Escherichia coli accounts for 70%–95% of urinary tract infections (UTIs). UTI is a serious health problem with respect to antibiotic resistance and biofilms formation being the prime cause for the antibiotic resistance. Biofilm can restrict the diffusion of substances and binding of antimicrobials. In this context, the present study is aimed to perform in vitro detection of biofilm formation among E. coli strains isolated from urine and to correlate their susceptibility pattern with biofilm formation. MATERIALS AND METHODS: A total of 100 E. coli strains isolated from patients suffering from UTI were included in the study. The identification of E. coli was performed by colony morphology, Gram staining, and standard biochemical tests. The detection of biofilm was carried out by Congo Red Agar (CRA) method, tube method (TM), and tissue culture plate (TCP) method. Antimicrobial sensitivity testing was performed by Kirby–Bauer disc diffusion method on Muller–Hinton agar plate. RESULTS: Of the 100 E. coli strains, 49 (49%) and 51 (51%) were from catheterized and noncatheterized patients, respectively. Biofilm production was positive by CRA, TM, and TCP method were 49 (49%), 55 (55%), and 69 (69%), respectively. Biofilm producers showed maximum resistance to co-trimoxazole (73.9%), gentamicin (94.2%), and imipenem (11.6%) when compared to nonbiofilm producers. Significant association was seen between resistance to antibiotic and biofilm formation with a P = 0.01 (<0.05). CONCLUSION: A greater understanding of biofilm detection in E. coli will help in the development of newer and more effective treatment. The detection of biofilm formation and antibiotic susceptibility pattern helps in choosing the correct antibiotic therapy.

Multidrug-resistant Acinetobacter septicemia in neonates: A study from a teaching hospital of Northern India Asifa Nazir Journal of Laboratory Physicians 2019 11(1):23-28 BACKGROUND: Acinetobacter species are typical nosocomial pathogens causing infections and high mortality, almost exclusively in compromised hospitalized patients. Acinetobacter sp. are intrinsically less susceptible to antibiotics and have propensity to acquire resistance. Multidrug-resistant (MDR) Acinetobacter sp. blood infection in the neonatal intensive care unit patients create a great problem in hospital settings. AIMS: A prospective data analysis was performed over a one year period of all neonates admitted with sepsis who developed Acinetobacter infection and their antibiotic susceptibility pattern was carried out. MATERIALS AND METHODS: Blood samples of infected neonates were collected aseptically and cases of Acinetobacter septicemia were identified. Speciation of Acinetobacter species was done. Various risk factors were identified and their drug-sensitivity test was performed. RESULTS: The incidence of neonatal septicemia due to Acinetobacter species was 13.7% (49/357). Predominant species isolated was Acinetobacter baumannii (98%). The major symptoms were lethargy and poor feeding. The major signs were tachypnea, intercostal retraction, and respiratory distress. The major fetal risk factors were low birth weight and prematurity. High degree of resistance was observed to the various antibiotics used. Majority of the isolates (95.9%) were MDR while 93.68% were resistant to carbapenems as well as extensively drug resistant. However, all the strains were sensitive to colistin. CONCLUSION: MDR Acinetobacter septicemia in neonatal patients is becoming alarmingly frequent and is associated with significant mortality and morbidity. Therefore, rational antibiotic use is mandatory along with an effective infection control policy in neonatal intensive care areas of each hospital to control Acinetobacter infection and improve outcome.

Volume, conductivity, and scatter parameters of leukocytes as early markers of sepsis and treatment response Parul Arora, Praveen Kumar Gupta, Raghavendra Lingaiah, Asok Kumar Mukhopadhyay Journal of Laboratory Physicians 2019 11(1):29-33 INTRODUCTION: Morphologic changes in the size and granularity of leukocytes seen in sepsis could be measured using the volume, conductivity, and scatter (VCS parameters) from the automated hematology analyzers. The objective of this study is to find the clinical usefulness of VCS parameters as possible indicators of sepsis and to determine the effect of treatment on these parameters. METHODS: This observational study was conducted in a tertiary level hospital in India. Hemogram and VCS parameters obtained from LH 750 (Beckman coulter, Fullerton, CA) from 134 proven blood culture-positive cases of sepsis were reviewed on the day of culture positivity (day 0), day 3, and day 7 were analyzed and compared with those of samples from otherwise healthy 100 participants. Statistical analysis of data was done, and cutoff value was established using receiver-operator characteristic curve. RESULTS: Out of 134 culture-positive cases, 55.2% (n = 74) Gram-negative and 44.8% (n = 60) Gram-positive bacteria were isolated. The mean neutrophil volume (MNV) and mean monocyte volume (MMV) were higher in the sepsis group compared to that of the control group (165.43 ± 18.21 vs. 140.59 ± 7.6, P = 0.001 for MNV and 179.8 ± 14.16 vs. 164.54 ± 9.6, P = 0.001 for MMV). A significant decrease in MNV and MMV was observed with the initiation of the treatment. Significant changes in scatter and conductivity parameters were also noticed. A cutoff value of 150.2 for MNV gave a sensitivity and specificity of 79.1% and 95%, respectively, with an area under the curve (AUC) of 92.3%. With a cutoff of 168.3, MMV had a sensitivity of 80.6% and specificity of 77.5%, AUC of 83%. CONCLUSION: VCS parameters such as MNV and MMV can be easily obtained by an automated hematology analyzer and could be used for early detection and therapeutic response in sepsis.

A 5-year surveillance on antimicrobial resistance of Acinetobacter isolates at a level-I trauma centre of India Minu Kumari, Priyam Batra, Rajesh Malhotra, Purva Mathur Journal of Laboratory Physicians 2019 11(1):34-38 INTRODUCTION: Acinetobacter spp. has emerged as a major cause of nosocomial outbreaks. Multiple antibiotic resistance is an important problem in Acinetobacter isolates in recent years. The aim of this study was to evaluate the rate of antimicrobial resistance and changes in resistance pattern over a period of 5 years (2012–2016) in Acinetobacter spp. isolated from trauma patients. MATERIALS AND METHODS: Acinetobacter spp. was identified by VITEK 2 and antibiotic susceptibility of isolates was investigated by disc-diffusion method and VITEK 2 automated system. Interpretation of susceptibility results was based on the Clinical and Laboratory Standards Institute guidelines. RESULTS: Out of the total 16,210 isolates obtained throughout the period of 5 years, Acinetobacter spp. accounted for 3744 (28.9%). Out of which, the species which was maximally isolated was Acinetobacter baumannii (98.5%), followed by Acinetobacter lwoffii (1.4%) and Acinetobacter hemolyticus (0.1%). The highest number of clinical isolates of Acinetobacter were recovered from neurosurgical ward (n = 1210), followed by the neurosurgical intensive care unit (ICU) (n = 1000) and surgical ICU (n = 948) and the most common sample of Acinetobacter isolation was from tracheal aspirate (37.1%), followed by wound swab (18.8%). The highest level of resistance was observed against ciprofloxacin (96%), followed by cefepime (95%), ceftazidime (95%), piperacillin (95%), and amikacin (92%). The trend of antibiotic resistance was found to be statistically significant (P < 0.001) for most of the antibiotics being tested such as amikacin and carbapenems. CONCLUSION: The high rate of antibiotic resistance of the Acinetobacter strains indicated that there is an urgent need for controlled antibiotic usage and appliance of hospital infection control measures.

Reconsidering azithromycin disc diffusion interpretive criteria for Salmonellae in view of azithromycin MIC creep among typhoidal and nontyphoidal salmonella Sadia Khan, Parvathy Kurup, Vivek Vinod, Raja Biswas, Gopala Krishna Pillai, Anil Kumar Journal of Laboratory Physicians 2019 11(1):39-44 PURPOSE: Enteric fever continues to be an important public health challenge for the developing world. With the emergence of fluoroquinolone resistance in Salmonellae spp. azithromycin is increasingly being used for oral treatment of enteric fever. We investigated the antibiotic susceptibility pattern of azithromycin in Salmonellae spp. isolates from a tertiary care hospital to detect emerging resistance. METHODS: The study assessed the reliability of disc diffusion as a screening test to detect azithromycin resistance by comparing it with the minimum inhibitory concentrations (MICs) of the drug in 100 Salmonellae spp. strains. The strains of Salmonellae spp. showing resistance to azithromycin were further investigated for resistance markers – mphA, mphB, and mef B genes. RESULTS: This study was conducted on 100 Salmonella enterica strains recovered from blood culture samples between 2013 and 2017. Among these isolates, 18 showed resistance to azithromycin by disc diffusion methodology with zones of inhibition <13 mm. MIC of 6 of these isolates were ≥32 mg/L. The mean MIC of azithromycin increased from 5 mg/L in 2013 to 24 mg/L in 2017. Azithromycin consumption as defined daily doses per 1000 patient days also showed an increase over the past 4 years. CONCLUSION: Azithromycin disc diffusion diameter interpretations as recommended by Clinical and Laboratory Standards Institute can mislabel a few sensitive strains as resistant. Azithromycin resistance is emerging in typhoidal and nontyphoidal Salmonella. MphA gene is associated with high MICs in nontyphoidal Salmonella spp.

Typing of Neisseria gonorrhoeae isolates by phenotypic and genotypic techniques in New Delhi, India Seema Sood, Neeraj Mahajan, Rajendra Singh, Sonu Kumari Agrawal, Trupti Shende, Arti Kapil, Hemanta K Kar, Vinod K Sharma Journal of Laboratory Physicians 2019 11(1):45-50 BACKGROUND: The objective of this study is to investigate gonococcal isolates using phenotypic and genotypic methods. METHODOLOGY: Sixty gonococcal isolates obtained were examined. Strains were divided into 9 resistant phenotypes: Chromosomally mediated penicillin-resistant Neisseria gonorrhoeae (CMRNGP), penicillinase-producing NG (PPNG), chromosomally mediated tetracycline-resistant NG (CMRNGT), TRNG, PPNG and TRNG, CMRNGPT, quinolone resistant NG (QRNG), Azithro R, and decreased susceptibility (DS) to ceftriaxone. These isolates were also subjected to auxotyping and NG-multi-antigen sequence typing (MAST). RESULTS: Of 60 isolates, 32 (53.33%) PPNG and only one was CMRNGP; 16 (26.66%) were CMRNGT, while 18 (30%) were TRNG. Both PPNG and TRNG found in 13 (21.66%) and none were CMRNGPT. QRNG was seen in 93.33%, 5% Azithromycin R, and 6.66% were DS to ceftriaxone. Based on auxotyping, 24 (40%) nonrequiring, 16 (26.66%) were proline requiring, 13 (21.66%) arginine requiring while 7 (11.66%) belonged to others. The most common ST was 6058 (32.5%). The discriminatory indices of antibiogram, auxotyping and NG-MAST were 0.77, 0.72, and 0.95, respectively. CONCLUSIONS: NG-MAST is the method of choice for epidemiological studies.

Correlation of vascular endothelial growth factor and vascular endothelial growth factor receptor-1 levels in serum and thyroid nodules with histopathological and radiological variables Gurkan Haytaoglu, Fatih Kuzu, Dilek Arpaci, Ayfer Altas, Murat Can, Figen Barut, Furuzan Kokturk, Sevil Uygun Ilikhan, Taner Bayraktaroglu Journal of Laboratory Physicians 2019 11(1):51-57 BACKGROUND/AIM: Vascular endothelial growth factor (VEGF) is a major cytokine in angiogenesis and has a role on aggressivity of various tumors. The expression of VEGF has been shown to increase in differential thyroid cancer. The aim of the study was to evaluate serum and intranodular VEGF (nVEGF) and VEGF receptor-1 (VEGFR-1) levels in patients with thyroid nodules and their relevance to ultrasonographic and pathological results. MATERIALS AND METHODS: A total of eighty patients were included in the study. Thyroid fine-needle aspiration biopsies were performed, and the levels of serum and nVEGF and VEGFR-1 were measured. Any possible correlations between serum and nVEGF, VEGFR-1, and biochemical/radiological variables were investigated. RESULTS: There were no significant differences between serum VEGF (sVEGF), nVEGF, sVEGFR-1, nVEGFR-1 levels, number of nodules, size of nodules, and benign and malignant ultrasonographic features. sVEGF and nVEGF were higher in malignant or suspicious nodules than that in benign nodules, but did not reach statistical significance (P > 0.05). sVEGFR-1 and nVEGFR-1 levels were higher in hyperthyroid patients than that in euthyroid patients (P < 0.05 and P = 0.003, respectively). nVEGFR-1 level was higher in hypothyroid patients than that in euthyroid patients (P = 0.016). sVEGF level was found to be higher in hyperactive nodules than that in others. Both sVEGFR-1 (P = 0.008) and nVEGF levels (P = 0.01) significantly increased with increasing age. nVEGFR-1 decreased with increasing body mass index (BMI) (P = 0.004). CONCLUSIONS: Our study showed the relationships of sVEGF, nVEGF, sVEGFR-1, and nVEGFR-1 levels with age, gender, BMI, and hyperthyroidism. To determine the role of VEGF/VEGFR-1 in thyroid nodules, further studies are required with a large number of patients.