Iranian Journal of Microbiology

Bioactivities of postbiotics in food applications: a review

2025-06-01T09:04:03+00:00

Postbiotics, which consist of beneficial compounds produced by probiotic bacteria, have emerged as promising natural pre- servatives in food applications. This article examines the health-promoting properties of postbiotics and their role in improv- ing food preservation and formulating nutrient-enriched foods. An organized investigation of published works was carried out through key research databases, including ScienceDirect, Scopus, PubMed, and Google Scholar, using keywords such as “Postbiotics,” “Biopreservation,” “Food Safety,” “Functional Foods,” “Antimicrobial Activity,” “Anti-inflammatory,” and “Bioactivities”. The findings reveal that postbiotics exert antimicrobial effects through multiple mechanisms, including the production of organic acids, bacteriocins, fatty acids, antimicrobial peptides, hydrogen peroxide, and vitamins. These bioac- tive substances actively suppress the proliferation of harmful and spoilage-causing microbes, consequently prolonging the preservation period of food items. Furthermore, postbiotics have been integrated into functional foods to modulate the host immune response and mitigate inflammatory conditions. Emerging applications of postbiotics also include their use in active food packaging systems, biofilm eradication, and cosmetic formulations. Although research on postbiotics is advancing, further investigations are required to elucidate the mechanisms of postbiotics and optimize their applications in both clinical and non-clinical contexts. This review emphasizes the potential of postbiotics to enhance food safety, improve nutritional quality, and contribute to overall health promotion.

Genotypic diversity of Mycobacterium tuberculosis strains collected from immigrant patients in Mashhad, Iran using MIRU-VNTR method

2025-06-01T09:04:02+00:00

Background and Objectives: This research aimed to explore the genetic diversity and phylogenetic relationships of My- cobacterium tuberculosis (Mtb) strains, as well as to assess their drug susceptibility, specifically in strains isolated from immigrant patients attending the Referral Tuberculosis Laboratory in Mashhad.

Materials and Methods: A total of 52 sputum samples isolated from patients were examined utilizing the Mycobacterial Interspersed Repetitive-Unit Variable Number of Tandem Repeats (MIRU-VNTR). Drug-susceptibility testing against ri- fampin (RIF) and isoniazid (INH) was measured utilizing the proportional strategy. Thereafter, for more examination, Xpert MTB/RIF and multiplex allele-specific PCR (MAS-PCR) was performed to determine RIF and INH-resistance within the Mtb strains.

Results: Among 52 Mtb isolates, 2 (3.8%) were resistant to rifampin and one isolate was resistant to both INH and RIF and considered as multidrug-resistance (MDR) isolate. According to MIRU-VNTR, the most prominent genetic-variation pat- terns of these samples, were related to NEW-1 (n=18, 34.6%), followed by CAS/Delhi (n=17, 32.7%), Haarlem (n=12, 23%), Uganda I (n=2, 3.8%), S (n=1, 1.9%), Beijing (n=1, 1.9%), and unknown (n=1, 1.9%) genotypes. The statistical analysis showed that the estimated percentage of the recent TB-transmission in this study was 0.21%.

Conclusion: The result of this study indicated a great diversity of MTBC circulating among Afghan-immigrants which might be one of the reasons for the infection to become active. The relatively high percentage of resistant isolates in the studied population shows the importance of screening the immigrants especially at the entry borders and treatment and follow up of patients, to control TB-incidence in country.

Phenotypic, genotypic characterization and antimicrobial resistance profiling of uropathogenic Escherichia coli in a tertiary care hospital, Puducherry, India

2025-06-01T09:04:00+00:00

Background and Objectives: Uropathogenic Escherichia coli (E. coli) (UPEC) accounts for 70-95% of community-ac- quired urinary tract infections (UTIs) and a significant proportion of nosocomial UTIs. This study aimed to characterize the phenotypic and genotypic characteristics of E. coli isolates from symptomatic UTI patients and evaluate their antimicrobial susceptibility patterns.

Materials and Methods: A hospital-based observational study was conducted at Aarupadai Veedu Medical College and Hospital, Puducherry, India, from August 2022 to April 2024. A total of 106 UPEC isolates were obtained from symptomatic UTI patients. Antimicrobial susceptibility testing (AST) was performed using the Kirby-Bauer method, and virulence genes (hlyA, fimH, papC) were detected using PCR.

Results: The mean age of patients was 49.7 years, with a female predominance (69.8%). Diabetes mellitus was the most common comorbidity (29.2%). Fever (60.4%) and dysuria (38.7%) were the most common symptoms. AST showed high susceptibility (>90%) to amikacin, nitrofurantoin, meropenem, and piperacillin/tazobactam, while >60% resistance was ob- served to cefotaxime and ceftazidime. Phenotypically, 30.2% of the isolates produced mannose-resistant hemagglutinins, and 17.9% produced hemolysin. ESBL production was found in 46.3%. Biofilm production was moderate in 65.1%, weak in 30.2% and strong in 4.7% and significantly correlated with multidrug resistance (p<0.05). Genotypically, 80.2% had fimH, 51.9% had papC and 20.8% had hlyA. papC was associated with reduced cefotaxime susceptibility (p<0.05).

Conclusion: The study highlights the significance of phenotypic and genotypic characterization in understanding UPEC virulence and resistance patterns, and emphasizes the need for targeted empiric therapy to improve UTI management.

Immunological and molecular detection of biofilm formation and antibiotic resistance genes of Pseudomonas aeruginosa isolated from urinary tract

2025-06-01T09:03:59+00:00

Background and Objectives: Pseudomonas aeruginosa (P. aeruginosa) is one of the most common causes of hospital-ac- quired infections. It is associated with high morbidity and healthcare costs, especially when appropriate antibiotic treatment is delayed. Antibiotic selection for patients with P. aeruginosa infections is challenging due to the bacteria's inherent resis- tance to many commercially available antibiotics. This study investigated antibiotic-resistance genes in isolated bacteria, which play a key role in disease pathogenesis.

Materials and Methods: 100 samples out of the 140 samples collected from urinary tract infections (UTIs) cases between December 15th, 2022, and April 15th, 2023, were included in the study. Identification of bacterial isolates was based on colony morphology, microscopic examination, biochemical tests, and the Vitek-2 system. Antibiotic resistance genes; Aph(3)-llla, ParC, Tet/tet(M), and aac(6´)-Ib-cr were tested by polymerase chain reaction (PCR).

Results: The obtained results were based on bacterial identifications of 81 clinical samples. Only 26 (32%) of these isolates were P. aeruginosa, 21 (26%) were Escherichia coli, and 18 (22.2%) were other bacteria. These isolates were used to detect four genes including tet(M), Aph(3)-llla, Par-c, and aac(6´)-Ib-cr. Four types of primers were used for PCR detection. The results showed that 11/14 (78.57%) carried the tet(M) gene, 10/14 (71.42%) carried the Aph(3)-llla gene, 14/14 (100%) car- ried the Par-c gene, and 10/14 (71.42%) of the isolates carried the aac(6´)-Ib-cr gene. The biofilm formation examining the esp gene, showed that 9 (64.28) isolates carried this gene.

Conclusion: The inability of antibiotics to penetrate biofilms is an important factor contributing to the antibiotic tolerance of bacterial biofilms.

Outpatient crystalluria: prevalence, crystal types, and associations with comorbidities and urinary tract infections at a provincial hospital

2025-06-01T09:03:58+00:00

Background and Objectives: Crystalluria refers to the occurrence of crystals in urine resulting from urinary supersatura- tion, which disrupts the balance between factors that promote and those that inhibit crystal formation in urine. This study aimed to assess the prevalence of crystalluria, identify crystal types, determine associated comorbidities, and assess links with bacterial urinary tract infections in outpatients at Hassan II Hospital in Settat.

Materials and Methods: A retrospective study was conducted from January 2022 to May 2023 at Hassan II Hospital. Urine samples from patients suspected of urinary tract infections, who underwent cytobacteriological urine examinations, were analyzed.

Results: Among 1,025 urine samples, 22.04% showed crystalluria. The mean age of patients was 51.3 with a standard de- viation of 18.1 years. The most common crystal types were calcium oxalate (46.4%), uric acid (23.5%), urates (15.1%) and struvite (9.3%). Comorbidities including, diabetes, kidney failure, prostatitis, and nephrotic syndrome was associated with urinary crystal formation. The prevalence of urinary tract infections in patients with urinary crystals was 10.6%. Struvite crys- tals were specifically associated with bacterial infections, especially with Proteus mirabilis, Escherichia coli, Citrobacter freundii, Citrobacter koseri, and Enterobacter cloacae.

Conclusion: Monitoring urinary crystals is essential for preventing the formation of kidney calculi and crystal-associated infections, especially in high-risk individuals.

Evaluation of the in vitro efficacy of antimicrobials against Enterobacterales with multiple carbapenemase enzymes

2025-06-01T09:03:57+00:00

Background and Objectives: High-dose of carbapenems and combination therapies with new β-lactam/β-lactamase inhib- itors and polymyxin B/tigecycline have been considered for treatment of carbapenem resistant Enterobacterales infection. The research was conducted to evaluate the in vitro potency of aminoglycosides, ceftazidime/avibactam/aztreonam and tigecycline against isolates of Enterobacteriaceae with multiple carbapenemase enzymes.

Materials and Methods: 42 genotypically confirmed carbapenem resistant Enterobacterales (twenty-nine NDM producers, nine NDM and OXA-48 producers, three NDM and VIM producers and one NDM combined with VIM and OXA 48 pro- ducer) were included. Minimum inhibitory concentration for carbapenems, aminoglycosides and tigecycline was determined by Vitek 2. Ceftazidime/avibactam/aztreonam synergy was observed by disk diffusion methodology.

Results: The in vitro efficacy of aminoglycosides was observed against Escherichia coli (E. coli) isolates with NDM and VIM genes. Low tigecycline susceptibility was observed among Klebsiella pneumoniae (K. pneumoniae) isolates with NDM and OXA-48 genes. Ceftazidime -avibactam/aztreonam combination displayed good in vitro activity against dual carbapene- mase producers of E. coli isolates (NDM with OXA-48 and NDM with VIM genes) and Klebsiella pneumoniae (combination of NDM, VIM and OXA-48 genes).

Conclusion: Ceftazidime/avibactam/aztreonam, aminoglycosides and tigecycline displayed in vitro activity against dual carbapenemase producers of E. coli and K. pneumoniae.

Prevalence of Helicobacter pylori infection among dyspeptic patients attending Baghdad medical city complex

2025-06-01T09:03:56+00:00

Background and Objectives: Dyspepsia is a disorder characterized by difficulty in digestion and represents a major health concern. Therefore, it is crucial to identify functional dyspepsia linked to Helicobacter pylori (H. pylori). This research aimed to determine the prevalence of H. pylori among patients with dyspepsia and to examine the potential risk factors associated with the infection.

Materials and Methods: From August 14th to September 21st, 2024, a total of 105 patients with dyspepsia, who attended the Central Laboratory of Baghdad Medical City Complex (Iraq), were enrolled in this study. Data on nonsteroidal anti-inflam- matory drugs (NSAIDs), smoking, family history, fasting habits and frequent fast food consumption were collected through participant interviews.

Results: Based on the urea breath test results, dyspeptic patients were categorized into infected (63.8%) and non-infected (36.2%) groups. Factors that influenced these patients included the intake of NSAIDs (48.6%), smoking (21.9%), family history (29.5%), fasting habits (36.2%) and regular consumption of fast food (57.1%).

Conclusion: Dyspeptic patients exhibit a high prevalence of H. pylori infection, indicating the significant impact of H. pylori on this population. However, the intake of NSAIDs, smoking, family history, fasting habits and regular fast food consump- tion have no significant effects on the presence of H. pylori.

Evaluation of the prevention, treatment and synergistic activity between Helicobacter pylori IgY antibodies and pantaprazole on AGS cell line and C57BL/6 Mice

2025-06-01T09:03:55+00:00

Background and Objectives: The growing resistance of Helicobacter pylori to antibiotics poses significant challenges in managing gastric ulcers. The application of IgY antibodies against H. pylori is a promising strategy. The current study evaluated the preventive and synergistic effects of IgY antibodies in conjunction with pantoprazole for treating H. pylori infection.

Materials and Methods: This investigation specifically focused on the inhibitory effects of IgY in the AGS cell line infected with H. pylori. In addition, the synergistic activity of IgY with pantoprazole and its preventive and therapeutic effects were assessed in male C57BL/6 mice.

Results: The findings indicated that IgY antibodies possess a substantial inhibitory effect on the adhesion of H. pylori to the AGS cell line. Furthermore, IgY antibodies resulted in a significant reduction (P<0.05) in the population of H. pylori and the severity of gastritis in infected C57BL/6 mice in both treatment and prevention groups. Notably, the optimal outcome was observed when IgY was administere d alongside pantoprazole.

Conclusion: The use of IgY has the potential to repair damaged cells and prevent infection by decreasing bacterial adherence to gastric epithelial cells. Given its synergistic effect with pantoprazole, IgY can be recommended as a suitable complemen- tary treatment in conjunction with pantoprazole.

Influence of Helicobacter pylori-derived outer membrane vesicles (OMVs) on Snail/β-Catenin cascade and metastasis-related proteins in 4T1 breast cancer cells

2025-06-01T09:03:54+00:00

Background and Objectives: This study investigates the impact of Helicobacter pylori (H. pylori)-derived outer membrane vesicles (OMVs) on the regulation of Snail/β-Catenin cascade and the production of metastasis-related proteins, such as E-cadherin and Vimentin, in the 4T1 cell line.

Materials and Methods: OMVs were purified from H. pylori (ATCC 700392) cultures and applied to 4T1 cells at concentra- tions of 1, 5, and 10 μg/mL, with untreated cells serving as controls. The MTT assay was employed to quantify cell viability. Expression profiles of +vimentin, Snail, α-SMA, and β-catenin genes were evaluated by qRT-PCR, while protein expression of E-cadherin and Vimentin was analyzed via immunohistochemistry. Data were analyzed using appropriate statistical meth- ods with SPSS and GraphPad Prism software.

Results: The MTT assay showed that 1 μg/mL OMVs were safe for normal cells. At this concentration, the expression of vimentin, Snail, α-SMA, and β-catenin genes significantly increased in the treatment group (P≤0.05). Additionally, Vimentin protein decreased, and E-cadherin protein increased (P≤0.05).

Conclusion: H. pylori-derived OMVs activate the Snail/β-Catenin cascade, influencing inflammatory responses and me- tastasis-related proteins, ultimately reducing migration in advanced cancer stages by modulating Vimentin and E-cadherin expression.

In vitro inhibition of Streptococcus mutans by probiotic yogurt fortified with Lactobacillus paracasei and Sargassum angustifolium protein hydrolysate: a functional yogurt for teeth decay prevention

2025-06-01T09:03:53+00:00

Background and Objectives: Probiotic yogurts enriched with Lactobacillus paracasei (L. paracasei), protein hydrolysates derived from Sargassum angustifolium macroalgae (SAPH), and encapsulated SAPH were formulated to inhibit Streptococ- cus mutans (S. mutans), the primary bacterium responsible for dental caries.

Materials and Methods: The yogurt samples were evaluated for physicochemical, microbiological, and sensory character-

istics.

Results: On day 21, the yogurt supplemented with L. paracasei demonstrated the greatest titratable acidity (97.35°D), the lowest pH value (4.24), reduced syneresis, and enhanced antioxidant, antihypertensive, and rheological properties. In terms of antibacterial activity, the lowest S. mutans count was detected in formulations containing free SAPH, either alone or in combination with L. paracasei. Conversely, yogurts formulated with encapsulated SAPH exhibited higher survival rates of both L. paracasei and S. mutans compared to those containing the free form of SAPH.

Conclusion: The findings indicated that although the probiotic yogurt containing free SAPH was more effective in reducing S. mutans levels within the yogurt matrix, the encapsulated form achieved an acceptable level of antibacterial activity while contributing to improved sensory acceptance.

Effects of lactic acid bacteria isolated from dairy products on lipid pattern of rats fed with a high fat diet

2025-06-01T09:03:51+00:00

Background and Objectives: A unique characteristic of probiotics is obesity and fatty liver control. In this study, the effect of lactic-acid-bacteria (LABs) isolated from dairy products was investigated on weight changes, blood biochemical indexes and liver tissue in mice fed a high-fat diet.

Materials and Methods: A total of 49 rats were assigned to 7 groups. The LAB-treated groups received the high-cholesterol diet supplemented with Lactobacillus reuteri, Lactobacillus plantarum, and Bifidobacterium animalis isolated from yogurt. At the end of 4 weeks, body weight changes, lipid factors and liver enzymes as well as liver lipid deposition and adipocyte size were measured.

Results: Serum low-density lipoprotein, total cholesterol, triglyceride levels and hepatic lipid deposition were significantly decreased in rats treated with LABs. The maximum and minimum weights were observed in the first and fourth weeks after treating with Lactobacillus and Bifidobacterium isolates, respectively. Liver enzymes were significantly decreased by LABs, especially in the group receiving concomitant administration of Lactobacillus and Bifidobacterium. Fatty liver process was reduced in the fat-fed group treated with L. reuteri.

Conclusion: LABs caused decreases in body weight gain, liver function, and adipocyte size. Therefore, coadministration of

Lactobacillus and Bifidobacterium in dairy products can significantly decrease lipid profile.

Anti-candida activity of Lacticaseibacillus rhamnosus R-2002 and its possible application in candidiasis prevention

2025-06-01T09:03:50+00:00

Background and Objectives: Probiotics are used for the treatment of yeast infections, they restore the balance in vaginal microbiome, adhere to epithelial cells, compete against pathogenic bacteria, acidify the environment, produce bacteriocins and modulate the immunity. The aim of the study was to investigate the anti-yeast activity (AYA) of the strain Lacticaseiba- cillus rhamnosus R-2002 against different Candida species.

Materials and Methods: From 20 strains of lactic acid bacteria examined, only L. rhamnosus R-2002 strain demonstrated beneficial properties against yeast. The effects of temperature and pH on AYA and its relation to cell wall were revealed by bi-layer agar assay. The connection of AYA to the cell wall was determined with the sonicated cells.

Results: R-2002 inhibited the growth of C. albicans ATCC 10291, C. tropicalis G 31 and C. albicans G4 (both isolated from vaginal samples). R-2002 maintained its AYA between a wide range of pH and its anti-yeast component/s are extracellular. The tested strain demonstrated stability against the high concentrations of progesterone and metronidazole, making it a suit- able candidate for the mitigation of vaginitis.

Conclusion: The present study summarizes all the positive features of the strain R-2002 and its potential as a therapeutic agent in the treatment of candidiasis.

In vitro effects of purified lacticin from whey isolated Lactococcus lactis culture on Staphylococcus aureus and MCF-7 breast cancer cells

2025-06-01T09:03:49+00:00

Background and Objectives: Bacteriocins are interested as antibacterial and anticancer agents due to their high specificity and low side effects. This study aimed to isolate bacteria which produce bacteriocins of the lacticin family from whey and to investigate their antibacterial and anticancer effects.

Materials and Methods: Lactic acid bacteria were isolated from different whey samples. The presence of the lacticin gene in the isolates was checked using PCR and then the inhibitory effects of their bacteriocin was investigated on Staphylococcus aureus utilizing well plate method. The protein content was separated by dialysis. The presence of lacticin was checked with the help of SDS-PAGE. The lacticin producing bacterium was identified through the sequencing of the 16S rRNA gene. Fi- nally, the cytotoxicity of the obtained protein was studied on the MCF-7 breast cancer cells using MTT and scratching tests.

Results: The isolated lacticin-producing Lactococcus lactis was able to grow in acidic conditions (pH = 2.5 for 3 h) and in bile salts (0.3% for 24 h). The bacterium produced 4.2 μg/μl bacteriocin with a molecular weight of 3.1 KD. The lacticin showed antibacterial effect against S. aureus. The cancerous cells treated with lacticin had slower growth than the controlin Scratch test. Based on the MTT results, more than 80% of cancerous cells were inhibited at a concentration of 7 μg/ml50 lacticin with IC= 5.2 μg/ml.

Conclusion: The bacteriocin produced in this study is a promising antibacterial and anticancer agent.

Exploring the antimicrobial potential of Rosmarinus officinalis against urinary tract infection isolates in Amman, Jordan

2025-06-01T09:04:04+00:00

Background and Objectives: The public health concern about urinary tract infections (UTIs) exists due to mounting an- tibiotic resistance rates. The antimicrobial properties of Rosmarinus officinalis create strong opportunities as an alternative therapeutic option. This study evaluated the antibacterial properties along with anti-biofilm behavior of rosemary extract against typical uropathogens.

Materials and Methods: This study collected samples from 500 UTI isolates for its cross-sectional research. The antibacte- rial activity of rosemary extract underwent testing for its effects on Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Pseudomonas aeruginosa through combination tests with disk diffusion, MIC and MBC assays. Biofilm inhi- bition was assessed using the Tissue Culture Plate method with extract concentrations of 25, 50, and 100 µg/mL. Statistical analysis included one-way ANOVA, Tukey's post-hoc, and regression analysis.

Results: The rosemary extract exhibited varying antibacterial effects, with inhibition zones ranging from 10 mm in E. faeca- lis to 16 mm in E. coli. MIC values were 4 mg/mL for E. coli and 32 mg/mL for E. faecalis, while MBC values ranged from 8 to 64 mg/mL. A 100 µg/mL concentration reduced E. coli biofilm formation by 70%. In checkerboard assays, rosemary extract enhanced antibiotic activity against E. coli and showed additive effects with K. pneumoniae and E. faecalis.

Conclusion: R. officinalis extract demonstrates promising antibacterial and anti-biofilm activities, suggesting potential as an adjunct UTI treatment, comparable to co-trimoxazole. Further research is recommended.

The inhibitory effect of the methanolic extract and the essence of Anvillea garcinii on expression of the genes related to Staphylococcus aureus biofilm formation

2025-06-01T09:03:48+00:00

Background and Objectives: Staphylococcus aureus (S. aureus) is a pathogenic bacterium whose virulence is attributed to its extracellular compounds and biofilm-forming ability. This study aimed to evaluate the inhibitory effects of the methanolic extract (AGME) and the essential oil (AGEO) of Anvillea garcinii on the growth and the biofilm formation of S. aureus.
Materials and Methods: The antibacterial and antibiofilm activities of AGME and AGEO against S. aureus ATCC 6538 were assessed using the microbroth dilution method and the Crystal Violet Staining Assay, respectively. The expression lev- els of sarA, spa, and icaA, genes involved in biofilm formation, were analyzed using real-time PCR.

Results: AGME and AGEO inhibited S. aureus growth at minimum inhibitory concentrations (MIC) of 1 mg/ml and 0.6 mg/ ml, respectively. AGME exhibited a 72% inhibition of biofilm formation at ¼ MIC, whereas AGEO showed no significant antibiofilm activity. AGME downregulated the expression of sarA, a key regulator of biofilm formation, as well as spa, and icaA genes.

Conclusion: This study demonstrated that A. garcinii essential oil (AGEO) exhibits significant antimicrobial activity, while its methanolic extract (AGME) effectively inhibits biofilm formation in S. aureus. These findings suggest the potential ap- plication of AGEO and AGME as antimicrobial and antibiofilm agents. Further investigations on their efficacy against other bacterial pathogens are recommended.

Diagnostic potential of new linear epitopes derived from the N-terminal domain of the SARS-CoV-2 Glycoprotein S

2025-06-01T09:03:46+00:00

Background and Objectives: The aim of this study was to assess the effectiveness of a new linear epitope from the N-ter minal domain (NTD) of the SARS-CoV-2 S protein in the diagnosis of COVID-19.

Materials and Methods: Serum samples from patients were confirmed to have COVID-19 by means of RT-PCR. The linear epitope sequence of the NTD was amplified by RT-PCR, inserted into an expression vector, and produced in Escherichi coli (DE3) pLysS. Subsequently, the recombinant proteins were purified and refolded. The interaction between the purified pro- tein and the antibodies in COVID-19 patient sera was evaluated using ELISA.

Results: Sequencing verified that the N-terminal linear epitope was successfully cloned into the PET-22b vector with a 6His-tag at the C-terminal end. The presence of a 25 kDa band on SDS-PAGE indicated the successful purification of the recombinant protein using Ni-NTA chromatography. The results of ELISA showed that the NTD linear epitope had strong sensitivity (88%) and specificity (96%) for identifying viral infection in COVID-19 patients' blood samples.

Conclusion: The findings of this study demonstrated that the NTD linear epitopes of the SARS-CoV-2 spike protein exhibit significant sensitivity and specificity for the diagnosis of COVID-19 infection using serological techniques. However, further evaluations involving larger sample sizes across diverse ethnic populations is essential.

Design and comparison of PCR-ELISA reaction with other available hybridization methods to identify types 11, 16, and 18 of the human papillomavirus

2025-06-01T09:03:46+00:00

Background and Objectives: Human papillomavirus (HPV) is a significant etiological agent in cervical cancer. This study aimed to evaluate the performance of PCR-ELISA for detecting HPV genotypes 11, 16, and 18 compared to the conventional hybridization methods.

Materials and Methods: PCR-ELISA was designed and optimized to detect target HPV genotypes using biotin-labeled probes. Sensitivity, specificity and reproducibility were assessed through intra-assay and inter-assay variability tests. Addi- tionally, a cost-benefit analysis was performed to compare PCR-ELISA with RT-PCR and gel electrophoresis.

Results: PCR-ELISA demonstrated high sensitivity (HPV18: 94.92%, HPV16: 98.36%, HPV11: 93.75%) and specificity (100% for all genotypes), with Kappa values ranging from 0.84 to 0.92, indicating strong agreement with the reference stan- dard. Reproducibility analysis showed intra-assay CVs below 5% for most samples and inter-assay CVs within acceptable limits. The cost-benefit analysis revealed significant reductions in reagent and equipment costs compared to RT-PCR, making PCR-ELISA a cost-effective alternative.

Conclusion: PCR-ELISA offers a reliable, sensitive and cost-effective method for HPV detection, particularly in re- source-limited settings. Its simplicity and compatibility with existing workflows makes it a promising tool for routine diag- nostic applications.

Optimized isolation and purification of native glycoprotein B from herpes simplex virus 1: a streamlined approach

2025-06-01T09:03:44+00:00

Background and Objectives: Viral membrane glycoproteins are essential for host cell recognition, membrane fusion and immune evasion, making them critical targets for antiviral therapies and vaccine development. However, their isolation in na- tive conformation is challenging due to structural complexity and limitations of conventional purification methods. The aim of current study was to develop a cost-effective, reproducible method for the isolation and purification of glycoprotein B (gB) from Herpes Simplex Virus type 1 (HSV-1) while maintaining its native conformation for functional and interaction studies. Materials and Methods: HSV-1 particles were concentrated via ultracentrifugation and membrane proteins were extracted using a modified protocol of the Mem-PER™ Plus Membrane Protein Extraction Kit. Native PAGE with a 4-8% gradient gel was employed to isolate multimeric gB (~300 kDa), followed by electroelution to extract the protein from the gel. The purity and integrity of gB were validated using SDS-PAGE and Western blot analysis.

Results: The method successfully isolated glycoprotein B in its native multimeric form with high purity and adequate con- centration (0.157 mg/mL). The pH of the native gel (8.3) and the high molecular weight of gB facilitated separation from other viral surface proteins. SDS-PAGE and Western blot confirmed the specificity and structural integrity of the purified protein.

Conclusion: This study introduces a cost-effective and reliable method for isolating viral glycoproteins in their native con- formation. The approach offers significant advantages over traditional chromatography-based techniques, making it ideal for research-scale applications, including functional and interaction studies.

Heavy hearts: the impact of influenza on young lives, a rare case report

2025-06-01T09:03:43+00:00

Considerable information is available about the acute respiratory symptoms of influenza A and B. However, rarely, these viruses can adversely affect the cardiovascular system. Few cases of pericardial effusion and cardiac tamponade due to the Influenza virus have been reported. To the best of our knowledge, we present the first case of a 23-year-old unvaccinated woman having concurrent influenza A and B infection manifesting as pericardial effusion and cardiac tamponade. The patient was treated with oseltamivir 75 mg, resulting in significant clinical improvement. This case emphasizes the importance of considering influenza as a possible cause of cardiac tamponade.