Medical Laboratory Journal

Challenges and diagnostic insights in extrapulmonary tuberculosis: A commentary on experiences from a high-burden region in Iran

Sadeghali Azimi

Editorial

Comparative evaluation of HPLC and nephelometry for HbA1c measurement: Diagnostic agreement and clinical utility

Battula Roopa Neeharika

Background: Glycated haemoglobin (HbA1c) reflects estimated average blood glucose levels over a period of 2-3 months and is essential for both diagnosing and managing diabetes. High-performance liquid chromatography (HPLC), particularly ion-exchange HPLC, is widely regarded as the reference and gold standard method for HbA1c assessment, whereas nephelometry is commonly adopted in clinical laboratories because it provides faster results. This study compared the accuracy, reliability, and clinical applicability of these two methods.
Methods: A total of 50 patients diagnosed with diabetes mellitus and attending a tertiary care hospital were included in this cross-sectional study. For each participant, HbA1c levels were estimated using both ion-exchange HPLC and nephelometric techniques. Data analysis involved descriptive statistics, Pearson’s correlation, the intraclass correlation coefficient (ICC), Bland-Altman plots, and receiver operating characteristic (ROC) curve evaluation. A paired t-test was performed to determine statistical significance, and values with p < 0.05 were considered statistically significant.
Results: The mean HbA1c values obtained by HPLC (6.2% ± 1.5) and nephelometry (6.3% ± 1.4) were closely aligned. A strong correlation was observed between the two techniques (r = 0.96, p < 0.01), and the intraclass correlation coefficient also indicated excellent concordance (ICC = 0.96). Bland-Altman plotting revealed only a slight bias, with a mean difference of 0.09%. ROC curve evaluation showed that both methods exhibited good diagnostic capability, although HPLC achieved marginally higher sensitivity (90%) and specificity (92%) than nephelometry (88% and 91%, respectively). While nephelometry offered faster processing, its diagnostic accuracy was slightly lower.
Conclusion: Although HPLC demonstrated superior diagnostic accuracy and method agreement, nephelometry offers operational advantages in high-volume settings. Despite the high correlation between the two methods, their interchangeability should be approached with caution, particularly in borderline cases. Further studies with larger and more diverse cohorts, as well as evaluation of potential confounding factors such as hemoglobin variants, are warranted.

Outbreak of Burkholderia gladioli septicemia in a neonatal intensive care unit: Investigation and control measures

Naila Begum

Background: Burkholderia gladioli (B. gladioli) is a rare but emerging pathogen associated with neonatal sepsis. This case series describes the investigation and control of an outbreak in a tertiary neonatal intensive care unit (NICU) in India.
Methods: In January 2024, seven neonates admitted to the NICU developed bloodstream infections. Clinical manifestations included respiratory distress, feeding refusal, metabolic acidosis, seizures, and shock. Predisposing factors included prematurity, congenital anomalies, and perinatal asphyxia. Blood cultures were processed using the automated BD BactecFX-40 system, and organism identification with antimicrobial susceptibility testing was performed using the BD Phoenix M-50 system. Environmental sampling was conducted to identify the outbreak source.
Results: Of 72 blood culture bottles, 34 (47.2%) flagged positive. B. gladioli were isolated from seven patients, accounting for 20.6% of positive cultures. The overall mortality rate was 57.1% (4/7 Deaths). All clinical isolates showed identical susceptibility patterns, with susceptibility to levofloxacin, cotrimoxazole, ceftazidime, chloramphenicol, minocycline, and meropenem, and intrinsic resistance to colistin. Environmental surveillance identified the same B. gladioli strain from suction apparatus and phototherapy units. Contributory factors included poor hand hygiene compliance (28-33%), limited glove availability, and overcrowding. Following cohorting, deep cleaning with 1% hypochlorite, staff retraining, and reinforcement of infection control protocols, no further isolates were recovered.
Conclusion: This outbreak highlights the high mortality associated with B. gladioli sepsis in neonates and underscores the critical need for stringent infection control practices and automated identification systems in NICUs. To our knowledge, this is the first such report from India.

Detection of the mecA gene and methicillin-resistant Staphylococcus aureus isolated from inanimate surfaces and healthcare workers in tertiary hospitals in Abia state, Nigeria

Ebubechi Uloma Okey-kalu

Background: Methicillin-resistant Staphylococcus aureus (MRSA) has become a public health issue. The purpose of this study was to detect MRSA among S. aureus isolates and to detect the presence of the mecA gene among selected MRSA isolates.
Methods: A total of 206 S. aureus isolates identified after the necessary biochemical tests were tested for methicillin resistance using the cefoxitin disk diffusion method. The Kirby-Bauer disk diffusion method was used to determine the antibiotic susceptibility pattern of the isolates. Some MRSA isolates were tested for the presence of the mecA gene.
Results: A total of 122 S. aureus isolates were recovered from inanimate surfaces, with footwear showing the highest contamination rate (39.34%, n = 48), while 84 S. aureus isolates were obtained from the palms of the hands and nostrils of healthcare workers, with the nostrils harboring the highest proportion (83.33%, n = 70). Antibiotic susceptibility testing was carried out on the S. aureus isolates. The S. aureus isolates were more resistant to ampicillin, erythromycin, cotrimoxazole, and gentamicin. A total of 206 S. aureus isolates were tested for MRSA using cefoxitin disk diffusion, and 43.69% (n = 90) were methicillin-resistant. A total of 20 S. aureus isolates suspected to be MRSA were tested for the mecA gene, and only 4 isolates (20% of the total isolates tested by polymerase chain reaction) were positive for the mecA gene.
Conclusion: The presence of MRSA in the two tertiary hospitals poses a risk to healthcare and community environments by contaminating healthcare workers, patients, visitors, and surrounding surfaces.

Discordant detection of anti-centromere antibodies by indirect immunofluorescence and ELISA: A case report and diagnostic considerations

Mohammadreza Sheikh Sajjadieh

Background: Anti-centromere antibodies (ACA) are commonly detected by indirect immunofluorescence (IIF) on HEp-2 cells, typically producing the classic centromere pattern (AC-3). However, discrepancies between IIF and solid-phase immunoassays, such as ELISA, may complicate interpretation.
Case Presentation: We report the case of a 75-year-old woman who presented with progressive inflammatory polyarthritis, morning stiffness, dry mouth, photosensitivity, and mild exertional dyspnea. Antinuclear antibody (ANA) testing by IIF on HEp-2 cells demonstrated a multiple nuclear dots pattern (AC-6), with features resembling a CENP-F–like pattern (AC-14), rather than the typical centromere pattern. By contrast, solid-phase assays, including ELISA and immuno-dot blot, demonstrated anti-centromere antibody reactivity. Additional serologic findings included strong positivity for SSA/Ro60, Ro52/TRIM21, SSB/La, and DFS70 antibodies.
Conclusion: This case demonstrates that anti-centromere reactivity detected by solid-phase assays may occur despite the absence of a classic AC-3 pattern on IIF. The interpretation of discordant serologic findings requires careful integration of clinical features with complementary laboratory methods.

Comparison of serum levels of zinc, copper, and selenium between patients with coronary artery disease and healthy controls: Selenium could be a new diagnostic biomarker

Reza Saghiri

Background: Although several clinical and laboratory diagnostic approaches have been developed for coronary artery disease (CAD), rates of mortality and morbidity remain high, possibly because of limitations in existing diagnostic markers. Here, we evaluated the serum levels of selected trace elements (Zn, Se, and Cu) in CAD patients and healthy subjects and assessed their diagnostic values in CAD.
Methods: 53 cardiologist-approved CAD patients and 48 age- and sex-matched healthy controls were recruited. Serum levels of Zn, Se, and Cu were assessed using the atomic absorption method. GraphPad v.8.4 and SPSS v.18 software were used for statistical analyses.
Results: Serum levels of Zn and Se were significantly lower in CAD patients than in controls (P = 0.0008 and P < 0.0001, respectively). In contrast, CAD patients showed significantly higher Cu levels (P = 0.0064). Regarding the ROC curve analyses, the area under the curve (AUC) for Zn was 0.6563 (P = 0.0069). Setting the cut-off value at 1.255 μg/mL yielded a sensitivity of 56.60%, specificity of 66.67%, and likelihood ratio (LR) of 1.698. The AUC for Se was 0.8595 (P<0.0001). The optimum cut-off value of 86.50 ng/ml yielded a sensitivity of 83.02%, specificity of 75.00%, and LR of 3.321. The AUC for Cu was 0.6557 (P = 0.0071). The optimum cut-off value of 1.225 μg/mL yielded a sensitivity of 62.26%, specificity of 64.58%, and LR of 1.758.
Conclusion: Our study showed that selenium may be a biomarker with reliable diagnostic value for CAD.

Comparison of clinical biochemistry indices between type 2 diabetic patients with nephropathy and non-nephropathy

Seyed Mostafa Mir

Background: Diabetic nephropathy is a major cause of kidney failure. The current study aimed to measure serum and urine albumin, as well as other biochemical markers, in individuals with type 2 diabetic nephropathy and to identify rapid diagnostic approaches in these patients.
Methods: This case-control study included 40 patients with diabetic nephropathy and 40 diabetic patients without nephropathy as the control group. Clinical parameters were assessed. After biochemical analysis and based on the ratio of random urine albumin to creatinine, patients were divided into two groups: those without albuminuria and those with microalbuminuria. SPSS version 18 software was used to analyze the results.
Results: In diabetic patients, the levels of urea, creatinine, and uric acid were significantly higher compared with the control group and were 64.68±46.84, 2.01±1.82, and 5.35±2.23, respectively. The average serum albumin content in diabetic patients with nephropathy was notably lower than that in patients without nephropathy (3.74±0.88 and 4.35±0.28, respectively) (p < 0.05). In patients with diabetic nephropathy, urinary albumin had a direct relationship with serum creatinine level (r=0.347, p=0.028). In the control group, serum albumin status could be assessed by measuring creatinine (r = -0.305, p = 0.056), urea (r = -0.333, p = 0.036), and HbA1C (r = -0.376, p = 0.017).
Conclusion: The results of this study revealed that high urinary albumin levels in patients with diabetic nephropathy have a direct association with serum creatinine levels, indicating a useful marker for evaluating nephropathy patients with albuminuria. However, further studies with larger numbers of participants are required.

Association between lipid metabolism markers and gastric cancer stage and grade: A Focus on ApoB

Hamid Reza Joshaghani

Background: Gastric cancer (GC), particularly adenocarcinoma, remains a major global health burden with high mortality, largely because of late-stage diagnosis and the limited availability of reliable biomarkers for disease monitoring. Lipid metabolism plays a crucial role in tumor biology, and serum lipid-related markers, including apolipoproteins, have been proposed as potential non-invasive indicators of tumor progression. This study aimed to evaluate the association between serum lipid profiles and tumor stage and histological grade in patients with gastric adenocarcinoma.
Methods: Fifty patients diagnosed with gastric adenocarcinoma were enrolled. Serum levels of total cholesterol, triglycerides, HDL-C, LDL-C, ApoA1, and ApoB were measured. Patients were categorized into early (Stages I-II) and advanced (Stages III-IV) tumor stages, as well as into moderately differentiated versus poorly differentiated grades. The Shapiro-Wilk test was used to assess data normality. Parametric and non-parametric tests were applied accordingly. A p-value < 0.05 was considered statistically significant.
Results: ApoB was the only parameter showing a significant association with tumor stage. Patients with advanced-stage GC had significantly lower mean ApoB levels than those in early stages (52.4 ± 2.6 vs. 63.4 ± 5.2 mg/dL, p = 0.042). No statistically significant differences were observed in ApoA1, HDL-C, total cholesterol, triglycerides, LDL-C, or VLDL with respect to either tumor stage or histological grade.
Conclusion: ApoB levels appear to decline with advancing tumor stage in gastric adenocarcinoma, suggesting a potential role as a marker for evaluating disease burden. Although no association was found with tumor grade, further validation in larger prospective studies incorporating metabolic and inflammatory covariates is warranted.

Investigation of Cryptosporidium oocyst contamination in water from Zabol and Zahedan: A molecular and microscopic analysis

Mansour Dabirzadeh

Background: Cryptosporidium is one of the most important protozoan parasites causing waterborne diseases worldwide. The parasite’s oocysts are resistant to conventional water treatment, making molecular detection crucial for identifying contamination sources.
Methods: A total of water samples were collected from different sites in Zabol and Zahedan, southeastern Iran. Microscopic screening was performed after concentration and staining with modified Ziehl-Neelsen and Trichrome methods under 1000× oil-immersion magnification. DNA was extracted from positive samples, and the SSU rRNA gene (~800-900 bp) was amplified by PCR. The resulting products were subjected to enzymatic digestion using AluI and RsaI restriction enzymes, and representative amplicons were sequenced.
Results: Microscopic examination confirmed the presence of Cryptosporidium oocysts in several water samples. PCR amplification successfully produced fragments of the expected size without nonspecific bands. AluI digestion revealed distinct fragment patterns consistent with Cryptosporidium spp., while RsaI showed no cutting sites. Sequence analysis through BLAST showed high identity (≥99%) with C. parvum isolates. The phylogenetic tree constructed using the BLAST distance-tree method grouped the sequence closely with C. parvum, confirming its identity.
Conclusion: Molecular characterization of Cryptosporidium from water samples in southeastern Iran indicated contamination primarily with C. parvum. These findings emphasize the necessity of continuous molecular surveillance to ensure the safety of drinking and recreational waters in the region.

Characterization and optimization of keratinase production by bacillus pumilus and bacillus tequilensis isolated from poultry waste

Fatemeh Haddadi

Background: The poultry industry produces a large amount of waste, including chicken feathers, which are difficult to decompose and can cause environmental pollution. Keratinase enzymes that degrade keratin may be used in poultry waste bioremediation. The objectives were to screen keratinolytic isolates, identify them, optimize culture conditions, and measure activity.
Methods: Two keratinolytic isolates were screened from poultry waste around Mashhad, Iran. They were identified, culture conditions were optimized, and activity was measured using azokeratin and turbidity absorbance.
Results: The two isolates were identified as Bacillus pumilus and Bacillus tequilensis. The optimal conditions were pH 7.0, 37°C, and 48 h of incubation. The maximum keratinase activity was 120 U/mL and 100 U/mL, respectively.
Conclusion: The two Bacillus isolates showed potential for poultry waste bioremediation and keratinase production applicable to animal feed and cleaning products.

Prevalence of fluoroquinolone resistance genes in the Escherichia coli sequence type 131 clone isolated from hospitalized patients with urinary tract infection

Mohammad Kargar

Background: The emergence of fluoroquinolone (FQ) resistance in the Escherichia coli (E. coli) sequence type 131 (ST131) clone has become a major challenge in the management of urinary tract infections (UTIs). Chromosomal mutations and plasmid-mediated quinolone resistance (PMQR) determinants play an important role in FQ resistance.
Methods: This cross-sectional study was conducted in 2020 on 300 urine samples. It aimed to investigate the prevalence of chromosomal mutations, PMQR genes, including qnr and aac (6′)-Ib-cr, and efflux pumps among FQ-resistant ST131 and non-ST131 E. coli causing UTIs. Initially, the ST131 clone was detected using allele-specific PCR and confirmed by Multilocus Sequence Typing.
Results: Among 95 FQ-resistant E. coli isolates, 29 (30%) belonged to the ST131 clone. The most frequently detected PMQR genes in FQ-resistant isolates were aac (6′)-Ib-cr and qnrS. However, statistical analysis revealed a stronger association between aac (6′)-Ib-cr and the ST131 clone (62%; p < 0.03). The oqxA gene was the most prevalent efflux pump gene observed in both ST131 (n = 11; 38%) and non-ST131 (n = 16; 24%) isolates. Analysis of the gyrA and parC genes revealed a significantly higher frequency of mutations in ST131 compared with non-ST131. Double mutations, S80I + E84V, were significantly more prevalent in both gyrA (76% ST131 vs. 43% non-ST131; p = 0.004) and parC (55% ST131 vs. 26% non-ST131; p = 0.002). Notably, the substitution E84G was exclusive to non-ST131 isolates (n = 4). High-level resistance (MIC ≥ 32 μg/mL) was observed in 96.6% (28/29) of ST131 isolates compared with 65% (43/66) of non-ST131 isolates.
Conclusion: The double mutations confer high-level resistance to FQs in the ST131 clone. These findings regarding resistance mechanisms can guide infection control strategies.