DNA Analysis Research Articles

Azolla pinnata and Lemna minor as comparative hyperaccumulators for livestock wastewater treatment: morpho-physiological and genetic approach.

The potential of two different aquatic macrophytes, Azolla pinnata R.Br. and Lemna minor L., to treat livestock wastewater through phytoremediation was investigated. The physiological analysis includes the removal efficiency of manganese (Mn) from livestock wastewater by AAS. Morphological observation was performed by using a scanning electron microscope (SEM) and visual observation. RAPD analysis was applied to observe the DNA profile. It was observed that the removal efficiency of Mn was higher in L. minor with a 92% removal rate, while in A. pinnata RE, it was at a 77% rate. The higher removal rate of Mn by L. minor showed that plants had a significant impact on the removal of heavy metals, with a p ≤ 0.05. Retention time and the removal of heavy metals were found to be positively correlated. As early as 24h after exposure to livestock wastewater (LW), the stomata on the leaves of A. pinnata and L. minor have both shrunk, and the root surfaces have shortened. According to the RAPD analysis, A. pinnata only shows an increase in band intensities and no polymorphism, whereas L. minor has 19% polymorphisms that indicate higher tolerance as hyperaccumulators. As a conclusion, L. minor showed no signs of necrosis and performed more efficiently as a hyperaccumulator in LW, with a higher removal efficiency.

DNA fingerprinting: a genetic database to give greater guarantees to the nursery chain

DNA fingerprinting: a genetic database to give greater guarantees to the nursery chain

Multiple ancestral components and complex origins of the Yugur people in Gansu province revealing by 35 Y-STR.

The Yugur people represent one of the ethnic groups residing within the Hexi Corridor, distinguishable by their small population size, linguistic diversity, intricate ancestral components, serving as a quintessential exemplar of the populations inhabiting this corridor. There are still many controversial issues in the academic community regarding the origin, migration, and formation process of the Yugur. In this study, we explored the formation process of the Yugur from the perspective of molecular anthropology, based on the paternal genetic characteristics of the Yugur people. And the study will synthesize multiple disciplines, encompassing ethnology, history and linguistics, in order to offer a thorough analysis. Within this research endeavor, a high-resolution kit comprising 35 Y-STRs was employed to examine 237 male specimens from the Yugur people in Gansu province. Y-SNP haplogroups were deduced through the utilization of Y-STR data. The paternal genetic data from diverse populations documented in published literature were merged to construct a 16 Y-STR dataset, a 25 Y-STR dataset, and a dataset detailing haplogroup frequencies. In this study, we employed haplotype network analysis, principal component analysis, multidimensional scaling analysis, phylogenetic tree construction, and genetic distance calculations to delve into the genetic structure, haplotype distribution, and genetic relationship with neighboring populations of the Yugur people. The findings of this study reveal that the Yugur people are a blend of ancestral lineages from both Eastern and Western Eurasian origins, with approximately 13% of their genetic component traced back to Western Eurasian populations. The Yugur people in Gansu province exhibits a more intimate genetic relationship with the Han, Tibetan, and Mongolian populations inhabiting nearby regions, while showing distinct genetic differences with Turkic-speaking groups like the Uyghur. Based on the merged data, we identified Q1b1a3-L330 and R1a1a-M17 shared with Turkic-speaking people, C2a1a1-F1756, C2a1a3-M504, C2a1a3a-F3796, C2a1a2-M48 and C2b1a1a1a-M407 shared with Mongolic-speaking people, D1a1a-M15 and D1a1b1-P47 shared with Tibetans, and multiple paternal lineages shared with Han people, which are the main paternal lineages of Yugur people, indicating multiple ancestral components and complex origins of Yugur. In this study, we provided a clearer genetic landscape which further supports the formation process and population characteristics of the Yugur people recorded in history, ethnology, and linguistics, and lays the foundation for more detailed studies on population genetics and forensic genetics in the future.

Molecular characterization of ST15 carbapenem-resistant Klebsiella pneumoniae isolated in a single patient

The carbapenem-resistant K. pneumoniae (CRKP) poses a serious threat to antibiotic applicability and public health. During treatment, K. pneumoniae (KP) frequently exhibits shifts in drug-resistant phenotypes, complicating clinical treatment as it transitions from sensitivity to resistance. In this study, we analyzed the clinical and molecular characteristics of drug resistance changes in KP strains isolated from a single patient, and the potential mechanisms underlying these resistance changes. Antimicrobial susceptibility test and string test were conducted to evaluate the resistant and virulent characterization of the strains. Pulsed field gel electrophoresis (PFGE) was used to investigate the homology relationship between the strains. The whole genome sequencing and phylogenetic analysis of 9 representative isolates was also performed. The transfer ability of the drug-resistant plasmid was studied by plasmid conjugation experiment. The transconjugants were verified by PCR amplification of specific genes, antimicrobial susceptibility test and PFGE. Our results revealed that 9 KP strains, isolated from the same patient, exhibited 'resistance-sensitivity-resistance-sensitivity' alternately to carbapenems. The differences in DNA fingerprint bands among the nine KP isolates were ≤ 3, which can be classified as the same PFGE type. Phylogenetic analysis showed that these 9 strains constituted a distinct branch within the phylogenetic tree. All nine KP strains belonged to the ST15-KL19 clone. Six of the strains were classified as CRKP, all of which carried eleven drug resistance genes: oqxB, oqxA, fosA6, aac(3)-lld, blaSHV-28, blaKPC-2, blaOXA-1, mph(A), tet(A), catB3 and aac(6')-lb-cr, mediating drug resistance to quinolones, fosfomycin, aminoglycosides, β-lactam, carbapenems, macrolides and chloramphenicol, belonging to multi-drug resistant bacteria. The carbapenem-resistant plasmid p2-KP3762-1 was found to transfer within species, from CRKP to hypervirulent K. pneumoniae (hvKP) RJF293HA, carbapenem-sensitive K. pneumoniae (CSKP) KP3657 and E. coli C600 at a frequency of (1.19±1.58) ×10-6, (1.09±1.38) ×10-7 and (10.9±9.53) ×10-6 respectively, resulting in the dissemination of carbapenem resistance genes. In this study, KP strains isolated from a single patient exhibited an alternating phenotype of resistant-sensitive-resistant-sensitive to carbapenems. The 9 KP isolates share a high degree of genetic similarity. The plasmid p2-KP3762-1, harboring the carbapenem resistance gene blaKPC-2, may undergo inter-strain and inter-clone transfer via conjugation in the patient during treatment. Furthermore, our findings suggest that the pathogens in this patient are likely to have a common ancestral origin.

The effect of commonly employed forensic DNA extraction protocols on ssDNA/dsDNA proportion and DNA integrity.

The effect of commonly employed forensic DNA extraction protocols on ssDNA/dsDNA proportion and DNA integrity.

Using an interaction timeline to investigate factors related to shedder status

A major factor that influences DNA transfer is the propensity of individuals to ‘shed’ DNA, commonly referred to as their ‘shedder status’. In this work we provide a novel method to analyse and interrogate DNA transfer data from a largely uncontrolled study that tracks the movements and actions of a group of individuals over the course of an hour. By setting up a model that provides a simplistic description of the world, parameters within the model that represent properties of interest can be iteratively refined until the model can sufficiently describe a set of final DNA observations. Because the model describing reality can be constructed and parametrised in any desired configuration, aspects that may be difficult to traditionally test together can be investigated. To that end, we use a 60- minute timeline of activity between four individuals and use DNA profiling results from objects taken at the conclusion of the hour to investigate factors that may affect shedder status. We simultaneously consider factors of: the amount of DNA transferred per contact, the rate of self-DNA regeneration, the capacity of hands to hold DNA, and the rate of non-self-DNA removal, all of which may ultimately contribute to someone’s shedder status.

Results of the 2023 Rapid DNA Multi-Laboratory Study – I-Chip

Identical sensitivity and mixture sample sets were provided to six external laboratories to evaluate the recent enhancements to the Rapid DNA technology for crime scene-type samples. Data were returned to the Federal Bureau of Investigation and the National Institute of Standards and Technology for review and analysis. Manual interpretation (modified Rapid DNA analysis) was conducted on all samples to assess the success of genotyping for both full profiles and the 20 Combined DNA Index System (CODIS) core STR loci using 18 total cellular DNA input amounts ranging from 102.4ng to 0.025ng. Two-person and three-person mixtures at varying ratios for two total DNA inputs were analyzed and observed allele percentages were compared to expected allele percentages.

Results of the 2023 Rapid DNA Multi-Laboratory Study – RapidINTEL Plus Sample Cartridge

Identical sensitivity and mixture sample sets were provided to six external laboratories to evaluate the recent enhancements to the Rapid DNA technology for crime scene-type samples. Data were returned to the Federal Bureau of Investigation and the National Institute of Standards and Technology for review and analysis. Manual interpretation (modified Rapid DNA analysis) was conducted on all samples to assess the success of genotyping for both full profiles and the 20 Combined DNA Index System (CODIS) core STR loci using 18 total cellular DNA input amounts ranging from 102.4ng to 0.025ng. Two-person and three-person mixtures at varying ratios for two total DNA inputs were analyzed and observed allele percentages were compared to expected allele percentages.

Impact of prescribed fire on fungal communities in Scots pine (Pinus sylvestris) forests in Mediterranean transitional zones

The Mediterranean region is renowned for its natural susceptibility to wildfires. In recent years, this risk has intensified due to various factors, including climate change and rural abandonment. Castilla y Leon stands out as one of the most severely impacted areas grappling with rural exodus. This evolving scenario accentuates the urgency of implementing forest management strategies to mitigate the escalating threat of wildfires, with a primary focus on fuel reduction. Although prescribed fires represent an efficient and cost-effective tool for wildfire prevention, they remain a contentious subject in Europe. Fungi not only contribute significantly to rural economies but also play a pivotal role in maintaining the equilibrium of forest ecosystems. The main objective of this study was to examine the effects of prescribed burning on Pinus sylvestris soils, with a specific focus on the recovery of fungal populations after such fires. To assess the short-term effects of prescribed fires on soil fungal communities, we collected soil samples from both burned and unburned plots 12 months post-burning to perform genomic DNA analyses. Our findings indicate that prescribed burning does not significantly alter fungal diversity or composition, with only litter saprotrophs showing significantly higher levels of abundance in burned areas than in unburned areas. Valuable edible fungi persisted post-burning, suggesting that prescribed burning could be used to reduce wildfire fuel loads while preserving fungal biodiversity and valuable edible fungi. These results advocate for the use of prescribed burning as a viable, myco-friendly forest management practice, offering a balance between fire prevention and ecological conservation.

Adrenocortical carcinoma with circulating tumor DNA analysis at post-operative recurrence: a case report with review of literature.

Recently, the usefulness of circulating tumor DNA (ctDNA) analysis in various malignancies has been reported. However, reports on ctDNA analysis in adrenocortical carcinoma (ACC) are few. Therefore, this study aimed to examine the detectability of genetic mutations in ctDNA and the association between ctDNA allelic ratio and disease progression in a patient with post-operative recurrence of ACC. A 77-year-old woman presented with a 5.4 cm left adrenal mass, which was clinically diagnosed as subclinical cortisol-producing ACC on close examination. She underwent left adrenalectomy and was diagnosed with stage II (T2N0M0) ACC. Post-operatively, adjuvant chemotherapy with mitotane was commenced because of histologically high-grade ACC. However, 17 months post-operatively, she had a local recurrence at the left adrenalectomy site. FoundationOne® CDx Cancer Genome Profile showed CTNNB1 G34A mutation in the resected adrenal tumor. She had heart failure and interstitial pneumonia and was treated with radiotherapy for local recurrence. Subsequently, lung and liver metastasis appeared post-operatively at 21 and 23 months, respectively. Serum dehydroepiandrosterone sulfate and computed tomography findings at 27 months post-operatively showed disease progression. We collected the peripheral blood at 23 and 27 months post-operatively and analyzed 18 genes associated with adrenal disease in plasma cell-free DNA and the resected adrenal tumor using a next-generation sequencer. At both time-points, CTNNB1 mutations consistent with the primary tumor were observed in ctDNA, with the allelic ratio increasing over time from 8% to 27%. In conclusion, monitoring the ctDNA allelic ratio may be useful for evaluating disease progression in advanced ACC.

Manipulation and analysis of large DNA molecules by controlling their dynamics using micro- and nano-gaps.

Manipulation and analysis methods for large DNAs are critical for epidemiological, clinical, diagnostic, and fundamental research on bacteria, membrane vesicles, plants, yeast, and human cells. However, the physical properties of large DNAs often challenge their manipulation and analysis with high accuracy and speed using the conventional methods such as gel electrophoresis and column-based methods. This review presents the approaches that leverage micrometer- and nanometer-sized gaps within microchannels to control the dynamics and conformations of large DNAs, thereby overcoming these challenges. By designing gap structures and migration conditions based on the relationship between gap parameters and the physical characteristics of large DNAs-such as diameter and persistence length-these methods enable swifter and more precise manipulation and analysis of large DNAs, including size separation, concentration, purification, and single-molecule analysis.

A Cross-Sectional Study of Variant Interpretation and Reporting of NGS Data Using Tertiary Analysis Software: Navify® Mutation Profiler.

Personalized medicine has revolutionized the clinical management of patients with solid tumors. However, the large volumes of molecular data derived from next-generation sequencing (NGS) and the lack of harmonized bioinformatics pipelines drastically impact the clinical management of patients with solid tumors. A possible solution to streamline the molecular interpretation and reporting of NGS data would be to adopt automated data analysis software. In this study, we tested the clinical efficiency of the Navify Mutation Profiler (nMP) software in improving the interpretation of NGS data analysis in diagnostic routine samples from patients with solid tumors. This study included one coordinating institution (Federico II University of Naples) and five other Italian institutions. Variant call format (VCF) files from reference standard samples previously tested by the coordinating institution and from n = 8 diagnostic routine samples (n = 2 from colorectal carcinoma; n = 2 from non-small cell lung cancer; n = 2 from advanced melanoma; and n = 2 from patients with gastrointestinal stromal tumors)and previously analyzed by each participating institution (n = 5) with standardized internal analysis workflows were uploaded onto the Navify® Mutation Profiler (nMP) system (Roche Sequencing Solutions, Pleasanton, CA, USA) for automated analysis and interpretation of DNA and RNA molecular alterations analytical parameters, molecular profiling, and clinical interpretation were carried out by the nMP system and compared with the standard workflow data analyzed by the participating institutions. Overall, all VCF files were successfully submitted and interpreted by the nMP system. A concordance agreement rate of 89.6% was observed between the automated and standard workflow systems. In particular, DNA and RNA molecular profiles obtained with the nMP system matched those obtained with standardized approaches in 44 out of 48 patients (91.7%) and in 11 out of 12 (91.7%) cases, respectively. In addition, the nMP system evidenced wild-type variants in 6 out of 7 (85.7%) cases. The nMP system represents a valid, easily manageable, and clinically useful system to interpret NGS data on diagnostic routine samples from patients with solid tumors.

Мультиплексный анализ микросателлитных маркеров в ДНК-паспортизации сортов черешни

Abstract. Microsatellite DNA markers are widely used in genetic studies of many fruit crops, including sweet cherry. They provide data that improve the efficiency of breeding and genetic research. Improving the methodological aspects of SSR genotyping is an important issue in order to reduce the costs of time, reagents and consumables while maintaining the accuracy of the analysis. One of these methods is the use of multiplex PCR, when several SSR loci are analyzed in one reaction. The aim of these studies was to improve the elements of the SSR fingerprinting methodology for sweet cherry varieties by developing multiplex sets of SSR markers. As a result of testing microsatellite DNA markers in the formation of multiplex sets, optimization of such parameters as the ratio of primer concentrations of DNA markers included in one multiplex set; the number of reaction cycles, the annealing temperature of primers during PCR; the degree of dilution of PCR products during their denaturation during fragment analysis on the ABIprism automatic genetic analyzer and Nanofor 05. Eight multiplex kits were developed, including 21 microsatellite DNA markers. Data on the polymorphism of DNA markers and on the length ranges of amplified fragments are presented. The obtained multiplex kits were successfully used for DNA certification of sweet cherry varieties representing the gene pool of the South of Russia. Part of the data on the DNA profiles of sweet cherry varieties was registered as an intellectual property object in the form of a database of SSR fingerprints of sweet cherry varieties. The obtained data can also be used to assess the genetic structure and diversity of collections of genetic resources of this crop. Keywords: sweet cherry, genotyping, SSR-markers, SSR-fingerprinting, multiplex analysis

Survey on forensic DNA biology training in forensic science service laboratories in the United States.

Training is an essential component of onboarding new hires in forensic science service provider (FSSP) laboratories. There are several DNA training standards published by the American Academy of Forensic Sciences (AAFS) Academy Standards Board (ASB) American National Standards Institute (ANSI) accredited framework. In this study, we conducted a survey of forensic DNA laboratory training programs to better understand training activities and materials. The survey was approved by the IRB and emailed to forensic laboratory directors, assistant directors, and/or DNA technical leaders and responses were submitted by them or their designees. Over thirty leaders and stakeholders responded. In this article, we report on the results of the survey. Respondents indicated that training activities included readings, writing assignments and quizzes, shadowing analysts, and mock casework and that training is documented and is a collaborative effort of the technical leader, unit supervisor, advanced forensic scientists, and other analysts and technicians. Laboratories assess competency using multiple methods including performance on mock casework, report writing, laboratory practical and competency tests, and a mock trial and testimony. The top three training activities reported are hands-on practice, shadowing, and readings. The top three focuses of the training are laboratory techniques (extraction, quantitation, amplification, and capillary electrophoresis), STR typing, and mixture analysis. Ethics violations and failure to pass the competency tests and mock trial, even after remediation, are the top reasons new hires fail training. Finally, the top items respondents would like to spend more time offering training on are troubleshooting, validation, and root cause analysis.

Forensic STR Loci and Schizophrenia: An Exploration of Implications for Forensic Applications and Genetic Privacy

Background/Objectives: Short tandem repeat (STR) loci are widely used in forensic genetics for identification and kinship analysis. Traditionally, these loci were selected to avoid medical associations, but recent studies suggest that loci such as TH01 and D16S539 may be linked to psychiatric conditions like schizophrenia. This study explores these potential associations and considers the privacy implications related to disease susceptibility. Methods: We analyzed 19 STR loci, including CODIS core loci and additional loci like Penta D and Penta E. Statistical analyses were conducted on a dataset of schizophrenia patients and matched control individuals to assess the relationship between STR polymorphisms and schizophrenia risk. Results: No significant associations were found between the 19 analyzed loci and schizophrenia in this dataset. While initial analyses revealed minor allele frequency differences at the D3S1358, D13S317, and TPOX loci between the schizophrenia and control groups, these differences did not retain statistical significance following Bonferroni correction (corrected p < 0.0026 for all loci). Conclusions: Although no significant associations were found between STR loci and schizophrenia, this study highlights the importance of considering the potential for forensic DNA data to reveal health-related information. As forensic DNA databases continue to expand, there is a growing need to reassess ethical and legal guidelines to ensure the protection of individual privacy. Future research should continue exploring these genetic associations with larger, more diverse samples to further understand their implications.

Discoveries by the genome profiling, symbolic powers of non-next generation sequencing methods.

Next-generation sequencing and other sequencing approaches have made significant progress in DNA analysis. However, there are indispensable advantages in the nonsequencing methods. They have their justifications such as being speedy, cost-effective, multi-applicable, and straightforward. Among the nonsequencing methods, the genome profiling method is worthy of reviewing because of its high potential. This article first reviews its basic properties, highlights the key concept of species identification dots (spiddos), and then summarizes its various applications.

Methanogens implicated by DNA evidence.

Methanogens implicated by DNA evidence.

ERBB2 comprehensive profiling and prognostication in Stage III Colon Cancer: Findings from PETACC8 and IDEA-France cohorts

ERBB2-pathway activation, through amplification or activating mutations, represents a new target for colon cancer (CC) treatment. We compared molecular methods to the gold-standard for assessing ERBB2 status and determined the prognostic value of ERBB2 amplification, mutations, and expression using data from two phase III trials involving nearly 3,000 stage III CC patients. In the PETACC8 trial, IHC/FISH, DNA, and RNA analysis were performed on 1813, 1719, and 1733 samples, respectively. In the IDEA-France trial, DNA and RNAseq were performed on 1129 and 1263 samples. The breast cancer SCAN-B cohort (n=3409) served as an external reference. A new molecular ERBB2-amplified status was defined using ERBB2 NGS-score, RNAseq expression, and clustering based on ERBB2 neighboring gene expression. Concordance between diagnostic techniques and the association between time-to-recurrence (TTR) and ERBB2-status were evaluated. The prevalence of the molecular ERBB2-amplified group was 1.85% in PETACC8 and 1.5% in IDEA-France, with a concordance of 0.81 (95% CI [0.70-0.92]) with the gold-standard IHC/FISH method in PETACC8. A non-linear relationship was observed between TTR and ERBB2-expression, with extreme groups showing a less favorable prognosis (P<.0001) in both colon and breast cancers. Patients with molecular ERBB2-amplified status or mutations had the poorest prognosis, followed by low-expression and intermediate-expression groups (3-year TTR 67.0%, 71.2%, and 77.9%, respectively). In multivariate analysis, the low-expression group had a significantly shorter TTR (HR=1.28; 95%CI [1.07-1.52]). The molecular definition of ERBB2-status could represent a cost-effective alternative in stage III CC. ERBB2 alterations and low RNA expression significantly reduce TTR, highlighting the complex role of ERBB2.

Genome-wide identification of SSR markers from coding regions for endangered Argania spinosa L. skeels and construction of SSR database: AsSSRdb.

Microsatellites [simple sequence repeats (SSRs)] are one of the most widely used sources of genetic markers, particularly prevalent in plants. Despite their importance in various applications, a comprehensive genome-wide identification of coding sequence (CDS)-associated SSR markers in the Argania spinosa L. genome has yet to be conducted. In this study, 66 280 CDSs containing 5351 SSRs within 4535 A. spinosa L. CDSs were identified. Among these, tri-nucleotide motifs (58.96%) were the most common, followed by hexa-nucleotide (15.71%) and di-nucleotide motifs (13.32%). The predominant SSR motif in the tri-nucleotide category was AAG (24.4%), while AG (94.1%) was the most abundant among di-nucleotide repeats. Furthermore, the extracted CDSs containing SSRs were subjected to functional annotation; 3396 CDSs (74.88%) exhibited homology with known proteins, 3341 CDSs (73.7%) were assigned Gene Ontology terms, 1004 CDSs were annotated with Enzyme Commission numbers, and 832 (18.3%) were annotated with KEGG pathways. A total of 3475 primer pairs were designed, out of which 3264 were successfully validated in silico against the A. spinosa L. genome, with 99.6% representing high-resolution markers yielding no more than three products. Additionally, the SSR markers demonstrated a low rate of transferability through in-silico verification in two species within the Sapotaceae family. Furthermore, we developed an online database, the "Argania spinosa L. SSR database: https://as-fmmdb.shinyapps.io/asssrdb/" (AsSSRdb) to provide access to the CDS-associated SSRs identified in this study. Overall, this research provides valuable marker resources for DNA fingerprinting, genetic studies, and molecular breeding in argan and related species. Database URL: https://as-fmmdb.shinyapps.io/asssrdb/.

Analysis of serotype distribution and characteristics of nonhemolytic and nonpigmented strains among group B Streptococcus isolates in a southern Taiwan local hospital.

Group B streptococci (GBS) are Gram-positive bacteria that are a leading cause of neonatal infections. Most invasive isolates are β-hemolytic, and hemolytic activity is critical for GBS virulence. Although nonhemolytic GBS strains are occasionally isolated, they are often thought to be attenuated in virulence. Recent studies have observed that many nonhemolytic and nonpigmented (NH/NP) strains originated from invasive infections, including bacteremia and meningitis, in neonates or adults. The mutations causing the NH/NP phenotype are predominantly localized in the cyl operon and abx1 gene. Previous studies on group B streptococci in Taiwan have focused on the serotype and genotype distribution. In this study, we investigated the serotype distribution of the NH/NP strains and detected the mutations of abx1. One hundred clinical GBS strains from non-invasive (vaginal and rectal swabs, 69) and invasive infections (blood, urine and abscess, 31), including 10 NH/NP isolates, were collected during 2019-2021 at Fooyin University Hospital. To confirm GBS isolates, we have developed a multiplex PCR method that detects GBS isolates, virulent strain ST-17 and virulent factor Srr1 simultaneously. Molecular serotyping was performed by multiplex PCR assay using serotype specific primer sets. The genomic region containing abx1 was amplified from DNA extracts by PCR and the amplicons were directly sequenced and analyzed on an ABI prism 3730 DNA analyzer. The capsular serotypes III and VI were the most abundant in both the non-invasive specimens and invasive specimens. The ST-17 isolates were more frequently associated with invasive infections (16.1%, 5/31) than non-invasive diseases or colonization (7.2%, 5/69). The association of NH/NP strains between noninvasive diseases or colonization (10.1, 7/69) and invasive infection (9.7%, 3/31) is nearly compatible. The NH/NP strains were isolated from various serotypes (Ia, III, V and VI) and five NH/NP isolates were serotype III. The virulence factor Srr1was detected in most of the NH/NP isolates (8/10) and one NH/NP isolate was ST-17. Abx1 mutations, including transitions, transversions and deletions, were observed in some NH/NP isolates, but some mutations also observed in hemolytic isolates. Five NH/NP isolates were erythromycin and clindamycin resistant. These results indicate NH/NP GBS strains may have the potential for invasive infections and may show higher tendency to get mutated.