Human metapneumovirus (HMPV) is a common respiratory pathogen that causes respiratory tract infections. In India, HMPV has been identified as one of the leading causes of morbidity and mortality in infants and young children with respiratory tract infections. The most reported sublineages of HMPV in India are B1, B2, A2b and A2c. A retrospective study was conducted to determine the circulating genotypes of HMPV among SARI cases from January 2016 to December 2018. Positive throat swab samples were confirmed with real-time RT-PCR. Subsequently, these samples were analysed using semi-nested conventional RT-PCR targeting the G gene, followed by sequencing and phylogenetic analysis. Clinical data analysis was also performed using SPSS 15.0 software. All 20 samples from the SARI cases were classified under the A2c sublineage of HMPV. Phylogenetic analysis indicated that these strains were genetically related to those circulating in Japan, China, and Croatia. Among the samples, ten showed 111-nucleotide duplications, while the other ten had 180-nucleotide duplications. Clinical analysis showed that four cases had coinfections with other pathogens. Our extensive analysis of patient samples determined that HMPV, especially the A2c genotype, significantly contributed to SARI cases within our study population, which signifies the importance of considering HMPV as a probable aetiological agent when investigating SARI outbreaks.

C1orf74, also known as URCL4, has been reported to have higher expression and be associated with poor prognosis in lung adenocarcinoma patients, and its role in regulation of the EGFR/AKT/mTORC1 pathway has been recently elucidated. In the current study, we used publicly available data and experimental validation of C1orf74 gene expression and its association with prognosis in cervical cancer patients. qRT-PCR was performed using RNA from cervical cancer cell lines and twenty-five cervical cancer patients. Data from TNMplot revealed that mRNA expression of the C1orf74 gene in primary tumor tissues, as well as metastatic tissues from cervical cancer patients, was significantly higher compared to normal cervical tissues. HPV-positive tumors had higher expression of this gene compared to HPV-negative tumors. qPCR analysis also demonstrated higher expression of C1orf74 in HPV-positive cervical cancer cell lines and most cervical cancer patients. The promoter methylation levels of the C1orf74 gene in cervical cancer tissues were lower compared to normal cervical tissues (p < 0.05). Collectively, our study indicates that higher expression of the C1orf74 gene caused by hypomethylation of its promoter is associated with poor overall survival in cervical cancer patients. Thus, C1orf74 is a novel prognostic marker in cervical cancer.

Human papillomavirus circulating free DNA (HPV cfDNA) is an emerging biomarker with potential utility in the detection and treatment monitoring of cervical cancer. To conduct a systematic review and meta-analysis evaluating the diagnostic and prognostic performance of HPV cfDNA in cervical cancer. A comprehensive literature search was conducted in PubMed, CINAHL, Cochrane Library, Scopus, and Embase through April 2025. Eligible studies reported or allowed calculation of diagnostic performance of HPV cfDNA in HPV-positive cervical cancer patients and/or included serial HPV cfDNA testing during post-treatment follow-up. Meta-analyses were conducted using a random-effects model. Heterogeneity was assessed with the I² statistic. The review followed PRISMA guidelines, and study quality was assessed using QUADAS-2. Of 106 studies screened, 20 met the inclusion criteria. Eleven studies contributed to the diagnostic meta-analysis and six to the prognostic analysis. The pooled sensitivity and specificity of HPV cfDNA for cervical cancer detection were 0.47 (95 % CI, 0.43-0.52) and 0.96 (95 % CI, 0.92-0.98), respectively. Positive and negative likelihood ratios were 10.49 and 0.28, with a diagnostic odds ratio of 71.31. The area under the SROC curve was 0.9825, indicating excellent overall diagnostic performance. Prognostically, HPV cfDNA positivity at 3 months post-treatment was significantly associated with reduced progression-free survival (HR = 8.50; 95 % CI, 4.69-15.41; I² = 0 %). HPV cfDNA shows high specificity and strong prognostic value, supporting its clinical utility in cervical cancer detection and treatment surveillance.

Human papillomaviral (HPV) integrations into host human genome, a key event in cervical carcinogenesis, are currently mapped through laborious and expensive sequencing methodologies. We developed and validated a novel library preparation strategy for nanopore sequencing to generate long targeted reads with HPV and human chimeric sequences. Using this strategy, we validated known HPV integrations in HeLa (HPV18) and SiHa (HPV16) cell lines. We also mapped integration sites in five HPV+ cervical cancer patients, which were confirmed by whole genome and Sanger sequencing. Our nanopore-based method provides a precise and efficient strategy to capture HPV integrations crucial for understanding tumorigenesis.

Detection of mutations conferring resistance to tyrosine kinase inhibitors (TKIs) in epidermal growth factor receptor (EGFR), such as T790M, is critical for guiding targeted therapies in non-small cell lung cancer (NSCLC) patients. However, limitations in accessing tumor tissue have given rise to liquid biopsy approaches necessitating ultrasensitive techniques to identify tumor mutations in a large background of wild-type alleles. We developed a sensitive and cost-effective technique utilizing a 2'3'-dideoxy nucleotide (ddNTP) blocker that selectively blocks wild-type allele amplification, allowing specific detection of mutant alleles in circulating free DNA (cfDNA) from plasma. EGFR genomic sequences encompassing T790M were cloned into pMD20-T plasmids and titrated in a background of genomic DNA to test the sensitivity. The assay detected 10 copies of the mutant allele in the presence of 10,000 copies of wild-type alleles, indicating a sensitivity of 0.001 %. Among 74 NSCLC patients, 34 received TKI treatment and were screened for the T790M mutation. The current method would be a sensitive and cost-effective liquid biopsy tool for detecting EGFR T790M mutations in NSCLC cfDNA samples, with strong potential for identifying actionable mutations for targeted therapy, especially in low-resource settings.