Displaying publications 41 - 60 of 100 in total

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  1. Guidelines and policies on collection of biological specimens in the Philippines. Philippine Congress, International Convention on Biodiversity
    Madulid DA
    J Ethnopharmacol, 1996 Apr;51(1-3):205-8.
    PMID: 9213618
    In October, 1993, 16 months after the United Nations approved the International Convention on Biodiversity held in Rio de Janeiro, June, 1992, the Philippine Congress ratified and adopted the Convention. This is a manifestation of the full support of the Philippines for the principles and policies adopted by the UN body on the conservation of biodiversity, sustainable development of biological resources and equitable sharing of benefits between users and owners of biodiversity resources. The Philippine scientific community has long recognized the need for and importance of a national guideline and policy with regard to the collection of plants and animals in the Philippines for scientific or commercial purposes. A series of consultative meetings were held by representatives of government agencies, non-government organizations, private organizations, academic and private persons concerned with biodiversity conservation to formulate national guidelines that regulate the collection of plant and animal specimens in the country. Guidelines were unanimously adopted by various government agencies and academia and a Memorandum of Agreement (MOA) was signed on September 28, 1990. Very recently a new document was drafted, specifically to serve as a guideline for those who desire to undertake sample collecting in the Philippines for biodiversity prospecting. The document is now being reviewed by government departments and agencies and will be presented to the President of the Philippines for signing as an Executive Order (EO). Once signed, this EO will serve as a national policy for bioprospecting in the country. The Philippines is one of the countries in Southeast Asia that has endorsed the adoption of regional guidelines on the collection of plant and animal organisms for drug development. The ASEAN Agreement on the Conservation of Nature and Natural Resources (1985). The Manila Declaration (1992) and lately, the Melaka Accord (1994), all of which were signed by various countries in Asia, are manifestations of this interest.
    Matched MeSH terms: Specimen Handling
  2. Tests on the centrifugal flotation technique and its use in estimating the prevalence of Toxocara in soil samples from urban and suburban areas of Malaysia
    Loh AG, Israf DA
    J Helminthol, 1998 Mar;72(1):39-42.
    PMID: 9639899
    The influence of soil texture (silt, sand and laterite) and flotation solutions (saturated NaCl, sucrose, NaNO3, and ZnSO4) upon the recovery of Toxocara ova from seeded soil samples with the centrifugal flotation technique was investigated. Soil samples of different texture were artificially seeded with Toxocara spp. ova and subjected to a centrifugal flotation technique which used various flotation solutions. The results showed significant (P < 0.001) interactions between the soil types and the flotation solutions. The highest percentage of ova recovery was obtained with silty soil (34.9-100.8%) with saturated NaCl as the flotation solution (45.3-100.8%). A combination of washing of soil samples with 0.1% Tween 80, and flotation using saturated NaCl and a 30 min coverslip recovery period was used to study the prevalence of contamination of soil samples. Forty-six soil samples were collected from up to 24 public parks/playgrounds in urban areas of Petaling Jaya and suburban areas of Serdang. The prevalence of Toxocara species in the urban and suburban areas was 54.5% and 45.8% respectively.
    Matched MeSH terms: Specimen Handling/methods
  3. Acceptability of Women Self-Sampling versus Clinician-Collected Samples for HPV DNA Testing: A Systematic Review
    Morgan K, Azzani M, Khaing SL, Wong YL, Su TT
    J Low Genit Tract Dis, 2019 Jul;23(3):193-199.
    PMID: 30933030 DOI: 10.1097/LGT.0000000000000476
    OBJECTIVES: Female self-sampling for human papillomavirus (HPV) DNA testing is an alternative screening method that can potentially increase cervical cancer screening coverage. This review addresses the acceptability of HPV DNA testing using self-sampling compared with conventional clinician-collected sampling. Barriers to and others factors associated with acceptability of either method were also examined.

    METHODS: The following electronic resources were searched: Medline @EBSCOHOST(Medline), Embase, PubMed, and CINAHL databases. Manual searches were also conducted. The main outcome of interest was the acceptability of HPV DNA testing by self-sampling in comparison with clinician-collected sampling.

    RESULTS: In total, 23 articles were included in this systematic review. The majority (19 studies) were quantitative intervention studies and 4 studies were qualitative observational studies. Eleven studies reported a preference for self-sampling by women compared with clinician-collected sampling (64.7%-93%). The remaining studies found that women preferred clinician-collected sampling because mainly of respondents' lack of confidence in their ability to complete self-sampling correctly. In most articles reviewed, the studied associated factors, such as demographic factors (age, marital status, and ethnicity), socioeconomic factors (income, education level), reproductive factors (condom use, number of children, current use of contraception, and number of partners), and habits (smoking status) were not found to be significantly associated with preference.

    CONCLUSIONS: Both methods of sampling were found to be acceptable to women. Self-sampling is cost-effective and could increase the screening coverage among underscreened populations. However, more information about the quality, reliability, and accuracy of self-sampling is needed to increase women's confidence about using to this method.

    Matched MeSH terms: Specimen Handling/methods*
  4. Development of a dry reagent-based triplex PCR for the detection of toxigenic and non-toxigenic Vibrio cholerae
    Chua AL, Elina HT, Lim BH, Yean CY, Ravichandran M, Lalitha P
    J Med Microbiol, 2011 Apr;60(Pt 4):481-485.
    PMID: 21183596 DOI: 10.1099/jmm.0.027433-0
    Vibrio cholerae has caused severe outbreaks of cholera worldwide with thousands of recorded deaths annually. Molecular diagnosis for cholera has become increasingly important for rapid detection of cholera as the conventional methods are time-consuming and labour intensive. However, traditional PCR tests still require cold-chain transportation and storage as well as trained personnel to perform, which makes them user-unfriendly. The aim of this study was to develop a thermostabilized triplex PCR test for cholera which is in a ready-to-use form and requires no cold chain. The PCR test specifically detects both toxigenic and non-toxigenic strains of V. cholerae based on the cholera toxin A (ctxA) and outer-membrane lipoprotein (lolB) genes. The thermostabilized triplex PCR also incorporates an internal amplification control that helps to check for PCR inhibitors in samples. PCR reagents and the specific primers were lyophilized into a pellet form in the presence of trehalose, which acts as an enzyme stabilizer. The triplex PCR was validated with 174 bacteria-spiked stool specimens and was found to be 100 % sensitive and specific. The stability of the thermostabilized PCR was evaluated using the Q10 method and it was found to be stable for approximately 7 months at 24 °C. The limit of detection of the thermostabilized triplex PCR assay was 2×10(4) c.f.u. at the bacterial cell level and 100 pg DNA at the genomic DNA level, comparable to conventional PCR methods. In conclusion, a rapid thermostabilized triplex PCR assay was developed for detecting toxigenic and non-toxigenic V. cholerae which requires minimal pipetting steps and is cold chain-free.
    Matched MeSH terms: Specimen Handling/methods
  5. Analyses of Epstein-Barr virus latent membrane protein-1 in Malaysian nasopharyngeal carcinoma: high prevalence of 30-bp deletion, Xho1 polymorphism and evidence of dual infections
    Tan EL, Peh SC, Sam CK
    J Med Virol, 2003 Feb;69(2):251-7.
    PMID: 12683415
    Nasopharyngeal carcinoma, a malignancy associated closely with Epstein-Barr virus (EBV), is prevalent among Chinese of Southern China origin. Epidemiological studies indicate a high prevalence of EBV in Asia with viral isolates having typical characteristics of the putative viral oncogene, latent membrane protein 1 (LMP-1), such as the loss of the Xho1 restriction site in Exon 1 and the 30-bp deletion in Exon 3. The EBV LMP-1 gene from throat washings of 120 nasopharyngeal carcinoma patients and 14 healthy individuals were analyzed. Similar analyses were also carried out on 30 and 12 postnasal space biopsies from nasopharyngeal carcinoma patients and healthy individuals, respectively. The 30-bp deletion was detected in 20% of nasopharyngeal carcinoma throat washes and in 100% of nasopharyngeal carcinoma postnasal space biopsies. Interestingly, 16% of the nasopharyngeal carcinoma biopsies possessed both the deleted and the undeleted variants, suggestive of dual infections. The notion of dual infections in nasopharyngeal carcinoma was further supported by the coexistence of both "F" and "f" (BamH1F region) EBV variants in 11% of the nasopharyngeal carcinoma biopsies. All of the throat washes and biopsies from the healthy controls showed the undeleted variant. The loss of the Xho1 restriction site was found with higher frequency both in throat washes and biopsies from patients with nasopharyngeal carcinoma. The discrepancy in the frequency of the 30-bp deletion between throat washes (20%) and postnasal space biopsies (100%) was an indication that this deletion is specific for viral isolates from primary tumour sites.
    Matched MeSH terms: Specimen Handling/methods
  6. Fulltext COVID-19 screening test by using random oropharyngeal saliva
    Rao M, Rashid FA, Sabri FSAH, Jamil NN, Seradja V, Abdullah NA, et al.
    J Med Virol, 2021 Apr;93(4):2461-2466.
    PMID: 33393672 DOI: 10.1002/jmv.26773
    An optimal clinical specimen for accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by minimizing the usage of consumables and reduce hazard exposure to healthcare workers is an urgent priority. The diagnostic performance of SARS-CoV-2 detection between healthcare worker-collected nasopharyngeal and oropharyngeal (NP + OP) swabs and patient performed self-collected random saliva was assessed. Paired NP + OP swabs and random saliva were collected and processed within 48 h of specimen collection from two cohort studies which recruited 562 asymptomatic adult candidates. Real-time reverse-transcription polymerase chain reaction targeting Open reading frame 1a (ORF1a) and nucleocapsid (N) genes was performed and the results were compared. Overall, 65 of 562 (28.1%) candidates tested positive for COVID-19 based on random saliva, NP + OP swabs, or both testing techniques. The detection rate of SARS-CoV-2 was higher in random saliva compared to NP + OP testing (92.3%; 60/65 vs. 73.8%; 48/65; p specimens. Our findings demonstrate that random saliva is an alternative diagnostic specimen for the detection of SARS-CoV-2. Self-collected random oropharyngeal saliva is a valuable specimen that provides accurate SARS-CoV-2 surveillance testing of a community.
    Matched MeSH terms: Specimen Handling/methods
  7. An improved stool concentration procedure for the detection of Cryptosporidium oocysts in Orang Asli stool samples
    Salleh FM, Moktar N, Yasin AM, Al-Mekhlafi HM, Anuar TS
    J Microbiol Methods, 2014 Nov;106:143-145.
    PMID: 25193442 DOI: 10.1016/j.mimet.2014.08.019
    To improve the stool concentration procedure, we modified different steps of the standard formalin-ether concentration technique and evaluated these modifications by examining stool samples collected in the field. Seven samples were found positive by the modified formalin-ether concentration technique (M-FECT). Therefore, the M-FECT procedure provides enhanced detection of Cryptosporidium oocysts.
    Matched MeSH terms: Specimen Handling/methods*
  8. Human Papilloma Virus (HPV) self-sampling: do women accept it?
    Abdullah NN, Daud S, Wang SM, Mahmud Z, Mohd Kornain NK, Al-Kubaisy W
    J Obstet Gynaecol, 2018 Apr;38(3):402-407.
    PMID: 29385850 DOI: 10.1080/01443615.2017.1379061
    This study aims to determine the acceptability of Human Papilloma Virus (HPV) self-sampling and the factors associated with willingness to buy HPV self-sampling kit in the future. A total of 164 women aged 28-60 years old from Obstetrics & Gynaecology clinics at a teaching hospital performed HPV self-sampling using the Digene HC2 DNA collection kit. After samples were taken, the participants were given self-administered questionnaires. The majority of the participants were Malay (93.9%), had attained tertiary education (65.2%) and were employed (70.1%). The acceptability was good. More than half of the participants felt that self-sampling was easy. Only 1.2% felt that the procedure was difficult to perform. Most reported no pain at all during the procedure (66.9%). The commonest concern was getting a good sample (90.1%). A number of Pap smears were found to be significantly associated with the willingness to buy the HPV self-sampling kit. HPV self-sampling has the potential to be included in the cervical cancer screening programme. Impact Statement What is already known on this subject: HPV self-sampling is acceptable in some developed and developing countries. It is acceptable because it was easy to perform with very minimal pain or discomfort. Studies on the acceptance of self-screening are needed to plan a policy on self-sampling in the future. What the results of this study add: Our study adds new findings to the body of knowledge on self-sampling in the local population. We found that more women are willing to do the self-sampling at the clinic rather than at home. Although more than 90% expressed willingness to do self-sampling in the future, only 70% of them were willing to purchase the kit. Cost is a potential barrier to women who have the interest to perform the self-sampling. Given the global economic challenges, cost is inevitably an important predictor that we have to consider. What the implications are of these findings for clinical practice and/or further research: Future research should examine women from the rural areas and those who are resilient to Pap smear screening. In clinical practice, clinicians should acknowledge that cost is a potential barrier for women who are willing to do self-sampling. Self-sampling could be an option for women with no financial constraint to buy the kit. However, clinicians should counsel women so that they can make an informed choice in determining their screening method.
    Matched MeSH terms: Specimen Handling/methods*
  9. Quantification of atovaquone from amorphous solid dispersion formulation using HPLC: An in vitro and ex vivo investigation
    Teoh XY, Goh CF, Aminu N, Chan SY
    J Pharm Biomed Anal, 2021 Jan 05;192:113631.
    PMID: 33011581 DOI: 10.1016/j.jpba.2020.113631
    Atovaquone (ATQ) is a poorly soluble drug. Therefore, formulating ATQ into its supersaturated state through solid dispersion for bioavailability enhancement can be of great value. However, due to fast crystallising properties of ATQ, the quantification of ATQ in a supersaturated solid dispersion system can be complicated. Therefore, in pursuit of accurate quantification of such sample, a simple HPLC analytical method utilising a C18 column (250 × 4.6 mm ID, 5 μm) for the quantitation of ATQ has been developed and validated. Atovaquone elution using the proposed method demonstrated a retention time around 7.6 min with good linearity (R2 > 0.999). The system suitability is also detailed with the tailing factor at 1.365 ± 0.002. The addition of solubilising agent as sample treatment step aided in ensuring the accurate quantitation of the fast crystallising ATQ. The developed HPLC quantitation method has been successfully employed in the analysis of ATQ from solid dispersion samples in in vitro dissolution as well as ex vivo permeation studies for formulation development.
    Matched MeSH terms: Specimen Handling
  10. Optimization of the storage conditions for coagulation screening tests
    Mohammed Saghir SA, Al-Hassan FM, Alsalahi OS, Abdul Manaf FS, Baqir HS
    J Coll Physicians Surg Pak, 2012 May;22(5):294-7.
    PMID: 22538033 DOI: 05.2012/JCPSP.294297
    To determine the optimum storage temperature and time for prothrombin time and activated partial thromboplastin time at various intervals at both room temperature and refrigerator.
    Matched MeSH terms: Specimen Handling
  11. Fulltext Advancing stem cell therapy from bench to bedside: lessons from drug therapies
    Srijaya TC, Ramasamy TS, Kasim NH
    J Transl Med, 2014;12:243.
    PMID: 25182194 DOI: 10.1186/s12967-014-0243-9
    The inadequacy of existing therapeutic tools together with the paucity of organ donors have always led medical researchers to innovate the current treatment methods or to discover new ways to cure disease. Emergence of cell-based therapies has provided a new framework through which it has given the human world a new hope. Though relatively a new concept, the pace of advancement clearly reveals the significant role that stem cells will ultimately play in the near future. However, there are numerous uncertainties that are prevailing against the present setting of clinical trials related to stem cells: like the best route of cell administration, appropriate dosage, duration and several other applications. A better knowledge of these factors can substantially improve the effectiveness of disease cure or organ repair using this latest therapeutic tool. From a certain perspective, it could be argued that by considering certain proven clinical concepts and experience from synthetic drug system, we could improve the overall efficacy of cell-based therapies. In the past, studies on synthetic drug therapies and their clinical trials have shown that all the aforementioned factors have critical ascendancy over its therapeutic outcomes. Therefore, based on the knowledge gained from synthetic drug delivery systems, we hypothesize that by employing many of the clinical approaches from synthetic drug therapies to this new regenerative therapeutic tool, the efficacy of stem cell-based therapies can also be improved.
    Matched MeSH terms: Specimen Handling/methods
  12. Detection of Epstein-Barr virus DNA in nasopharyngeal carcinoma using a non-radioactive digoxigenin-labelled probe
    Permeen AM, Sam CK, Pathmanathan R, Prasad U, Wolf H
    J Virol Methods, 1990 Mar;27(3):261-7.
    PMID: 2157729
    The presence of Epstein Barr virus (EBV) DNA in biopsies from the post-nasal space (PNS) of patients suspected of nasopharyngeal carcinoma (NPC) was detected by in situ cytohybridization with an EBV DNA probe labelled with the novel labelling compound digoxigenin. The digoxigenin probe was hybridised to cryostat sections of NPC biopsies and subsequently detected by an enzyme immunoassay procedure. It was found that in situ cytohybridization using the digoxigenin probe was much more rapid and sensitive (96 h compared to five weeks) than the current method of using 3H-labelled probe. Using the digoxigenin EBV probe, it was found that in all the eighteen NPC biopsies tested, EBV DNA was detected in malignant epithelial cells and infiltrating lymphocytes. EBV DNA was also detected in some normal epithelial cells in these NPC biopsies. EBV DNA was not detected in epithelial cells of non-malignant biopsies.
    Matched MeSH terms: Specimen Handling
  13. Fulltext Kehadiran Acanthamoeba spp. pada bekas penyimpanan kanta sentuh pengguna asimptomatik
    Putri Noradyani Megat Hashim, Mohamed Kamel Abdul Ghani, Norhayati Moktar, Anisah Nordin, Yusof Suboh, Mimi Fazah Zainudin, et al.
    Jurnal Sains Kesihatan Malaysia, 2007;5(2):53-61.
    MyJurnal
    Keratitis Acanthamoeba merupakan sejenis inflamasi kornea yang dikaitkan dengan penggunaan kanta sentuh. la disebabkan oleh Acanthamoeba spp., ameba hidup bebas yang tersebar luas di pelbagai persekitaran manusia. Kontaminasi Acanthamoeba spp. pada bekas penyimpanan kanta sentuh merupakan faktor kehadiran ameba pada kanta seterusnya menjangkiti mata. Kajian ini bertujuan untuk melihat kehadiran Acanthamoeba spp. pada bekas penyimpanan kanta sentuh pengguna asimptomatik. Seramai 90 orang pengguna kanta sentuh asimptomatik terlibat dalam kajian ini. Sampel diambil secara swab pada bekas kanta sentuh dan dikulturkan ke atas agar bukan nutrien yang dilapisi Escherichia coli. Plat agar diperiksa setiap hart bagi mengesan kehadiran ameba. Kultur positif seterusnya disahkan di bawah 'Image Analysis with Video TesT 4.0'. Acanthamoeba spp. didapati positif pada lapan daripada 90 sampel (8. 7%) dan kesemua strain adalah kumpulan II (polyphagids). Penemuan ini membuktikan Acanthamoeba spp. boleh Nadir pada bekas penyimpanan kanta sentuh pengguna asimptomatik dan boleh menjadi risiko jangkitan keratitis Acanthamoeba.
    Matched MeSH terms: Specimen Handling
  14. Fulltext Evaluation of blood sampling times and indications for therapeutic drug monitoring services
    Aishah Hamzah, Ab Fatah Ab Rahman
    Malaysian Journal of Pharmaceutical Science, 2008;6(1):1-11.
    MyJurnal
    The appropriateness of sampling times and indications for monitoring of serum drug concentrations for the purpose of therapeutic drug monitoring (TDM) were evaluated at three hospitals on the east coast of Malaysia. Appropriateness criteria for indication and sampling were adapted from previously published criteria and with input from local TDM pharmacists. Six drugs were chosen, namely gentamicin, digoxin, carbamazepine, phenobarbital, phenytoin, and valproic acid. A total of 265 TDM requests were evaluated. Appropriateness of the indication for TDM ranged from 77.4% to 82%, while that for sampling ranged from 34.2% to 62.1%. There were no significant differences between the three hospitals in both categories of appropriateness. Among different drug groups, the percentage of appropriate indication was found to be highest with antiepileptic drugs. Antiepileptic drugs, however, had the lowest rate of appropriate sampling. Overall, findings from the three hospitals showed very encouraging results with almost 80% of the requests considered as appropriately indicated. However, the percentage of appropriateness of sampling was lower, and thus may require further investigation.
    Matched MeSH terms: Specimen Handling
  15. Increasing Reproducibility in Oral Microbiome Research
    Gopinath D, Menon RK
    Methods Mol Biol, 2021;2327:1-15.
    PMID: 34410636 DOI: 10.1007/978-1-0716-1518-8_1
    Evidence on the role of the oral microbiome in health and disease is changing the way we understand, diagnose, and treat ailments. Numerous studies on diseases affecting the oral cavity have revealed a large amount of data that is invaluable for the advancements in diagnosing and treating these diseases. However, the clinical translation of most of these exploratory data is stalled by variable methodology between studies and non-uniform reporting of the data.Understanding the key areas that are gateways to bias in microbiome studies is imperative to overcome this challenge faced by oral microbiome research. Bias can be multifactorial and may be introduced in a microbiome research study during the formulation of the study design, sample collection and storage, or the sample processing protocols before sequencing. This chapter summarizes the recommendations from literature to eliminate bias in the microbiome research studies and to ensure the reproducibility of the microbiome research data.
    Matched MeSH terms: Specimen Handling
  16. A novel approach for collecting samples from fruit bats for isolation of infectious agents
    Chua KB
    Microbes Infect., 2003 May;5(6):487-90.
    PMID: 12758277
    During the outbreak of Nipah virus encephalitis involving pigs and humans in peninsular Malaysia in 1998/1999, a conventional approach was initially undertaken to collect specimens from fruit bats by mist-netting and shooting, as an integral part of wildlife surveillance of the natural reservoir host of Nipah virus. This study describes a novel method of collecting fruit bats' urine samples using plastic sheets for isolation of Nipah virus. This novel approach resulted in the isolation of several other known and unidentified infectious agents besides Nipah virus.
    Matched MeSH terms: Specimen Handling/methods*
  17. Fulltext Antibacterial Properties and Effects of Fruit Chilling and Extract Storage on Antioxidant Activity, Total Phenolic and Anthocyanin Content of Four Date Palm (Phoenix dactylifera) Cultivars
    Samad MA, Hashim SH, Simarani K, Yaacob JS
    Molecules, 2016 Mar 26;21(4):419.
    PMID: 27023514 DOI: 10.3390/molecules21040419
    Phoenix dactylifera or date palm fruits are reported to contain natural compounds that exhibit antioxidant and antibacterial properties. This research aimed to study the effect of fruit chilling at 4 °C for 8 weeks, extract storage at -20 °C for 5 weeks, and extraction solvents (methanol or acetone) on total phenolic content (TPC), antioxidant activity and antibacterial properties of Saudi Arabian P. dactylifera cv Mabroom, Safawi and Ajwa, as well as Iranian P. dactylifera cv Mariami. The storage stability of total anthocyanin content (TAC) was also evaluated, before and after storing the extracts at -20 °C and 4 °C respectively, for 5 weeks. Mariami had the highest TAC (3.18 ± 1.40 mg cyd 3-glu/100 g DW) while Mabroom had the lowest TAC (0.54 ± 0.15 mg cyd 3-glu/100 g DW). The TAC of all extracts increased after storage. The chilling of date palm fruits for 8 weeks prior to solvent extraction elevated the TPC of all date fruit extracts, except for methanolic extracts of Mabroom and Mariami. All IC50 values of all cultivars decreased after the fruit chilling treatment. Methanol was a better solvent compared to acetone for the extraction of phenolic compounds in dates. The TPC of all cultivars extracts decreased after 5 weeks of extract storage. IC50 values of all cultivars extracts increased after extract storage except for the methanolic extracts of Safawi and Ajwa. Different cultivars exhibited different antibacterial properties. Only the methanolic extract of Ajwa exhibited antibacterial activity against all four bacteria tested: Staphylococcus aureus, Bacillus cereus, Serratia marcescens and Escherichia coli. These results could be useful to the nutraceutical and pharmaceutical industries in the development of natural compound-based products.
    Matched MeSH terms: Specimen Handling
  18. Intestinal carriage of multidrug-resistant gram-negative bacteria in preterm-infants during hospitalization in neonatal intensive care unit (NICU)
    Yap PS, Ahmad Kamar A, Chong CW, Yap IK, Thong KL, Choo YM, et al.
    Pathog Glob Health, 2016 Sep;110(6):238-246.
    PMID: 27650884
    The prevalence and antibiotic susceptibility of intestinal carriage of Gram-negative bacteria among preterm infants admitted to the neonatal intensive care unit (NICU) in a tertiary teaching hospital in Malaysia were determined. A total of 34 stool specimens were obtained from preterm infants upon admission and once weekly up to two weeks during hospitalization. The presumptive colonies of Escherichia coli and Klebsiella pneumoniae were selected for identification, antibiotic susceptibility testing, and subtyping by using pulsed-field gel electrophoresis (PFGE). Out of 76 Gram-negative isolates, highest resistance was detected for amoxicillin/clavulanate (30.8%, n = 16), ceftriaxone (42.3%, n = 22), ceftazidime (28.8%, n = 15), cefoxitin (28.8%, n = 15), aztreonam (36.5%, n = 19), and polymyxin B (23.1%, n = 12). Three colistin resistant K. pneumoniae have also been detected based on E-test analysis. Thirty-nine isolates of K. pneumoniae and 20 isolates of E. coli were resistant to more than three antimicrobial classes and were categorized as multidrug resistant (MDR). PFGE analysis revealed a higher diversity in pulsotypes for K. pneumoniae (18 pulsotypes) in comparison to E. coli (four pulsotypes). In addition, a total of fifteen pulsotypes was observed from 39 MDR K. pneumoniae. The risk factors for antibiotic resistance were assessed using random forest analysis. Gender was found to be the most important predictor for colistin resistant while length, OFC, and delivery mode were showing greater predictive power in the polymyxin B resistance. This study revealed worrying prevalence rates of intestinal carriage of multidrug-resistant K. pneumoniae and E. coli of hospitalized preterm infants in Malaysia, particularly high resistance to polymyxins.
    Matched MeSH terms: Specimen Handling/methods
  19. Fulltext A novel strategy for community screening of SARS-CoV-2 (COVID-19): Sample pooling method
    Lim KL, Johari NA, Wong ST, Khaw LT, Tan BK, Chan KK, et al.
    PLoS One, 2020;15(8):e0238417.
    PMID: 32857823 DOI: 10.1371/journal.pone.0238417
    The rapid global spread of the coronavirus disease (COVID-19) has inflicted significant health and socioeconomic burden on affected countries. As positive cases continued to rise in Malaysia, public health laboratories experienced an overwhelming demand for COVID-19 screening. The confirmation of positive cases of COVID-19 has solely been based on the detection of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) using real-time reverse transcription polymerase chain reaction (qRT-PCR). In efforts to increase the cost-effectiveness and efficiency of COVID-19 screening, we evaluated the feasibility of pooling clinical Nasopharyngeal/Oropharyngeal (NP/OP) swab specimens during nucleic acid extraction without a reduction in sensitivity of qRT-PCR. Pools of 10 specimens were extracted and subsequently tested by qRT-PCR according to the WHO-Charité protocol. We demonstrated that the sample pooling method showed no loss of sensitivity. The effectiveness of the pooled testing strategy was evaluated on both retrospective and prospective samples, and the results showed a similar detection sensitivity compared to testing individual sample alone. This study demonstrates the feasibility of using a pooled testing strategy to increase testing capacity and conserve resources, especially when there is a high demand for disease testing.
    Matched MeSH terms: Specimen Handling/methods*
  20. Fulltext Assessment of Telomere Length in Archived Formalin-Fixed, Paraffinized Human Tissue Is Confounded by Chronological Age and Storage Duration
    Kong PL, Looi LM, Lau TP, Cheah PL
    PLoS One, 2016;11(9):e0161720.
    PMID: 27598341 DOI: 10.1371/journal.pone.0161720
    Telomeres shorten with physiological aging but undergo substantial restoration during cancer immortalization. Increasingly, cancer studies utilize the archive of formalin-fixed, paraffin-embedded (FFPE) tissues in diagnostic pathology departments. Conceptually, such studies would be confounded by physiological telomere attrition and loss of DNA integrity from prolonged tissue storage. Our study aimed to investigate these two confounding factors. 145 FFPE tissues of surgically-resected, non-diseased appendixes were retrieved from our pathology archive, from years 2008 to 2014. Cases from 2013 to 2014 were categorized by patient chronological age (0-20 years, 21-40 years, 41-60 years, > 60 years). Telomere lengths of age categories were depicted by telomere/chromosome 2 centromere intensity ratio (TCR) revealed by quantitative fluorescence in situ hybridization. Material from individuals aged 0-20 years from years 2013/2014, 2011/2012, 2009/2010, and 2008 were compared for storage effect. Telomere integrity was assessed by telomere fluorescence intensity (TFI). Epithelial TCRs (mean ± SD) for the respective age groups were 4.84 ± 2.08, 3.64 ± 1.21, 2.03 ± 0.37, and 1.93 ± 0.45, whereas corresponding stromal TCRs were 5.16 ± 2.55, 3.84 ± 1.36, 2.49 ± 1.20, and 2.93 ± 1.24. A trend of inverse correlation with age in both epithelial and stromal tissues is supported by r = -0.69, p < 0.001 and r = -0.42, p < 0.001 respectively. Epithelial TFIs (mean ± SD) of years 2013/2014, 2011/2012, 2009/2010 and 2008 were 852.60 ± 432.46, 353.04 ± 127.12, 209.24 ± 55.57 and 429.22 ± 188.75 respectively. Generally, TFIs reduced with storage duration (r = -0.42, p < 0.001). Our findings agree that age-related telomere attrition occurs in normal somatic tissues, and suggest that an age-based reference can be established for telomere studies on FFPE tissues. We also showed that FFPE tissues archived beyond 2 years are suboptimal for telomere analysis.
    Matched MeSH terms: Specimen Handling
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