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Increased sensitivity of malaria detection by nested polymerase chain reaction using simple sampling and DNA extraction

Cox-Singh J, Mahayet S, Abdullah MS, Singh B

Int J Parasitol, 1997 Dec;27(12):1575-7.
PMID: 9467744

Malaria remains a disease of underdeveloped and remote regions of the world. The application of polymerase chain reaction (PCR) technology to malaria epidemiology has the potential for increasing our knowledge and understanding of this disease. In order to study malaria in all geographical locations it is important that specimen collection and DNA extraction for PCR be kept simple. Here we report a method for extracting DNA from dried blood spots on filter paper which is capable of detecting one Plasmodium falciparum and two Plasmodium vivax parasites/microliter of whole blood by nested PCR without compromising the simplicity of specimen collection or DNA extraction.

Matched MeSH terms: Specimen Handling
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
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
Fulltext Intraspecific macroscopic digestive anatomy of ring-tailed lemurs (Lemur catta), including a comparison of frozen and formalin-stored specimens

Clauss M, Trümpler J, Ackermans NL, Kitchener AC, Hantke G, Stagegaard J, et al.

Primates, 2021 Mar;62(2):431-441.
PMID: 33180215 DOI: 10.1007/s10329-020-00873-8

Digestive tract measurements are often considered species specific, but little information exists on the degree to which they change during ontogeny within a species. Additionally, access to anatomical material from nondomestic species is often limited, with fixed tissues possibly representing the only available source, though the degree to which this material is representative in terms of dimensions and weight is debatable. In the present study, the macroscopic anatomy of the digestive tract (length of intestinal sections, and tissue weights of stomach and intestines) of 58 Lemur catta [ranging in age from 1 month (neonates) to 25 years], which had been stored frozen (n = 27) or fixed in formalin (n = 31), was quantified. Particular attention was paid to the caecum and the possible presence of an appendix. The intraspecific allometric scaling of body mass (BM)0.46[0.40;0.51] for total intestine length and BM0.48[0.41;0.54] for small intestine length was higher than the expected geometric scaling of BM0.33, and similar to that reported in the literature for interspecific scaling. This difference in scaling is usually explained by the hypothesis that, to maintain optimal absorption, the diameter of the intestinal tube cannot increase geometrically. Therefore, geometric volume gain of increasing body mass is accommodated for by more-than-geometric length scaling. According to the literature, not all L. catta have an appendix. No appendix was found in the specimens in the present study. The proportions of length measurements did not change markedly during ontogeny, indicating that the proportions of the foetus are representative of those of the adult animal. By contrast, width and tissue-mass scaling of the caecum indicated disproportionate growth of this organ during ontogeny that was not reflected in its length. Compared to overall intraspecific variation, the method of storage (frozen vs. formalin) had no relevant impact on length or weight measurements.

Matched MeSH terms: Specimen Handling/methods*
Fulltext Investigation of CPD and HMDS sample preparation techniques for cervical cells in developing computer-aided screening system based on FE-SEM/EDX

Jusman Y, Ng SC, Abu Osman NA

ScientificWorldJournal, 2014;2014:289817.
PMID: 25610902 DOI: 10.1155/2014/289817

This paper investigated the effects of critical-point drying (CPD) and hexamethyldisilazane (HMDS) sample preparation techniques for cervical cells on field emission scanning electron microscopy and energy dispersive X-ray (FE-SEM/EDX). We investigated the visualization of cervical cell image and elemental distribution on the cervical cell for two techniques of sample preparation. Using FE-SEM/EDX, the cervical cell images are captured and the cell element compositions are extracted for both sample preparation techniques. Cervical cell image quality, elemental composition, and processing time are considered for comparison of performances. Qualitatively, FE-SEM image based on HMDS preparation technique has better image quality than CPD technique in terms of degree of spread cell on the specimen and morphologic signs of cell deteriorations (i.e., existence of plate and pellet drying artifacts and membrane blebs). Quantitatively, with mapping and line scanning EDX analysis, carbon and oxygen element compositions in HMDS technique were higher than the CPD technique in terms of weight percentages. The HMDS technique has shorter processing time than the CPD technique. The results indicate that FE-SEM imaging, elemental composition, and processing time for sample preparation with the HMDS technique were better than CPD technique for cervical cell preparation technique for developing computer-aided screening system.

Matched MeSH terms: Specimen Handling/methods*
Fulltext Isolation of Acanthamoeba sp. from conjunctival sac of healthy individuals using swab

Anisah N, Amal H, Kamel AG, Yusof S, Noraina AR, Norhayati M

Trop Biomed, 2005 Jun;22(1):11-4.
PMID: 16880749 MyJurnal

Is Acanthamoeba sp. normally found in the eyes? A study was carried out to establish the possibility of Acanthamoeba sp. as a part of the normal conjunctival flora. Conjunctiva swabbing were carried out in 286 healthy Orang Asli school children using sterile cotton swab. The swab was then inoculated onto non-nutrient agar (NN-A). Heat killed Escherichia coli that was used as food source for the growth of the amoebae was pipetted onto and away from the smear. The plates were incubated at 30 degrees C and examined daily using an inverted microscope for 14 days. Morphology of the trophozoites and cysts of the amoebae were used as the taxonomic criteria for identification. Positive-controls and negative-controls were done to check for the consistency of the technique used and monitoring of contamination respectively. None of the conjunctiva swab cultured was positive for Acanthamoeba sp. This finding may indicate that Acanthamoeba sp. is not part of normal conjunctival flora or conjunctiva swab is an insensitive technique to isolate the organism. However, a more extensive research is needed to investigate these possibilities.

Matched MeSH terms: Specimen Handling/instrumentation
Fulltext Isolation of Janthinobacterium lividum from early onset neonatal sepsis patients in Malaysia

Shinkafi SH, Umar S, Neela VK, Noordin SM, Noordin SA, Hudu SA, et al.

Afr Health Sci, 2019 Sep;19(3):2378-2389.
PMID: 32127808 DOI: 10.4314/ahs.v19i3.11

Background: The term early onset neonatal septicaemia (EONS) refers to invasive bacterial infections that primarily involve the blood stream of neonates during the first 3 days of life. Although early onset neonatal septicaemia is relatively uncommon, it may be associated with case fatality rates of 15-30% and substantial morbidity in surviving infants.
Objectives: This study describes an unusual septicaemia cases with Janthinobacterium lividum in neonatal Intensive Care Units.
Methods: Bacterial causes of early onset neonatal sepsis in Kuala Lumpur Hospital Malaysia were investigated using broad range 16S rDNA PCR and sequencing. The bacterial DNA was isolated directly from blood without pre-incubation. All samples collected were equally cultured and incubated in automated BACTEC system.
Results: Two hundred and fifty two neonates were recruited in this study with mean (SD) gestational age of 35.9. Neonates with J. lividum infection lacked microbiological evidence of septicaemia as their blood culture yielded no bacterial growth. However, the PCR analysis of these samples yielded 1100bp corresponding to bacteria species.
Conclusion: This study demonstrates the value of PCR in detecting bacteria where special growth requirement is involved.

Matched MeSH terms: Specimen Handling
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
Fulltext Light microscopy and molecular identification of Sarcocystis spp. in meat producing animals in Selangor, Malaysia

Latif B, Kannan Kutty M, Muslim A, Hussaini J, Omar E, Heo CC, et al.

Trop Biomed, 2015 Sep;32(3):444-52.
PMID: 26695204 MyJurnal

One thousand and forty-five tissue samples of skeletal muscles, tongue, heart, diaphragm and esophagus were collected from 209 animals (43 sheep, 89 goats and 77 cattle) from an abattoir in Selangor between February and October, 2013. Each sample was divided into three pieces with each piece measuring 2-3 mm3. Each piece was then squeezed between two glass slides and examined microscopically at x 10 magnification for the presence of sarcocystosis. Three positive samples from each animal species were then fixed in 10% formalin for histological processing. Seven positive samples collected from each animal species were preserved at -80°C or 90% ethanol for gene expression studies. Microsarcocysts were detected in 114 (54.5%) animals by light microscopy (LM). The infection rates in sheep, goat and cattle were 86, 61.8 and 28.6% respectively. The highest rate of infection was in the skeletal muscles of sheep (64.9%) and goats (63.6%) and in the heart of cattle (63.6%). The cysts were spindle to oval in shape and two stages were recognized, the peripheral metrocytes and centrally located banana-shaped bradyzoites. 18S rRNA gene expression studies confirmed the isolates from the sheep as S. ovicanis, goats as S. capracanis and cattle as S. bovicanis. This, to the best of our knowledge, is the first molecular identification of an isolate of S. ovicanis and S. capracanis in Malaysia. Further studies with electron microscopy (EM) are required in the future to compare the features of different types of Sarcocysts spp.

Matched MeSH terms: Specimen Handling/methods
Fulltext Metagenomic data on bacterial diversity profiling of high-microbial-abundance tropical marine sponges Aaptos aaptos and Xestospongia muta from waters off terengganu, South China Sea

Amelia TSM, Lau NS, Amirul AA, Bhubalan K

Data Brief, 2020 Aug;31:105971.
PMID: 32685631 DOI: 10.1016/j.dib.2020.105971

Marine sponges are acknowledged as a bacterial hotspot and resource of novel natural products or genetic material with industrial or commercial potential. However, sponge-associated bacteria are difficult to be cultivated and the production of their desirable metabolites is inadequate in terms of rate and quantity, yet bioinformatics and metagenomics tools are steadily progressing. Bacterial diversity profiles of high-microbial-abundance wild tropical marine sponges Aaptos aaptos and Xestospongia muta were obtained by sample collection at Pulau Bidong and Pulau Redang islands, 16S rRNA amplicon sequencing on Illumina HiSeq2500 platform (250 bp paired-end) and metagenomics analysis using Ribosomal Database Project (RDP) classifier. Raw sequencing data in fastq format and relative abundance histograms of the dominant 10 species are available in the public repository Discover Mendeley Data (http://dx.doi.org/10.17632/zrcks5s8xp). Filtered sequencing data of operational taxonomic unit (OTU) with chimera removed is available in NCBI accession numbers from MT464469 to MT465036.

Matched MeSH terms: Specimen Handling
Fulltext Molecular identification of Malaysian Chrysomya megacephala (Fabricius) and Chrysomya rufifacies (Macquart) using life stage specific mitochondrial DNA

Kavitha R, Tan TC, Lee HL, Nazni WA, Sofian AM

Trop Biomed, 2013 Jun;30(2):211-9.
PMID: 23959486 MyJurnal

DNA identification of blow fly species can be a very useful tool in forensic entomology. One of the potential benefits that mitochondrial DNA (mtDNA) has offered in the field of forensic entomology is species determination. Conventional identification methods have limitations for sibling and closely related species of blow fly and stage and quality of the specimen used. This could be overcome by DNA-based identification methods using mitochondrial DNA which does not demand intact or undamaged specimens. Mitochondrial DNA is usually isolated from whole blow fly and legs. Alternate sources for mitochondrial DNA isolation namely, egg, larva, puparium and empty puparium were explored in this study. The sequence of DNA obtained for each sample for every life cycle stage was 100% identical for a particular species, indicating that the egg, 1st instar, 2nd instar, 3rd instar, pupa, empty puparium and adult from the same species and obtained from same generation will exhibit similar DNA sequences. The present study also highlighted the usefulness of collecting all life cycle stages of blow fly during crime scene investigation with proper preservation and subsequent molecular analysis. Molecular identification provides a strong basis for species identification and will prove an invaluable contribution to forensic entomology as an investigative tool in Malaysia.

Matched MeSH terms: Specimen Handling/methods
Non-Invasive DNA Sampling for Molecular Analysis of Beta-Thalassemia: Amiable Alternative Sampling Methods with Accurate Results for Pediatric Patients

Abd Rahim MR, Kho SL, Kuppusamy UR, Tan JA

Clin. Lab., 2015;61(9):1325-30.
PMID: 26554253

BACKGROUND: Beta-thalassemia is the most common genetic disorder in Malaysia. Confirmation of the β-globin gene mutations involved in thalassemia is usually carried out by molecular analysis of DNA extracted from leukocytes in whole blood. Molecular analysis is generally carried out when affected children are around 1 - 2 years as clinical symptoms are expressed during this period. Blood taking at this age can be distressing for the child. High yield and pure DNA extracted from non-invasive sampling methods can serve as alternative samples in molecular studies for genetic diseases especially in pediatric cases.
METHODS: In this study, mouthwash, saliva, and buccal cytobrush samples were collected from β-thalassemia major patients who had previously been characterized using DNA extracted from peripheral blood. DNA was extracted from mouthwash, saliva, and buccal cytobrush samples using the conventional inexpensive phenol-chloroform method and was measured by spectrophotometry for yield and purity. Molecular characterization of β-globin gene mutations was carried out using the amplification refractory mutation system (ARMS).
RESULTS: DNA extracted from mouthwash, saliva, and buccal cytobrush samples produced high concentration and pure DNA. The purified DNA was successfully amplified using ARMS. Results of the β-globin gene mutations using DNA from the three non-invasive samples were in 100% concordance with results from DNA extracted from peripheral blood.
CONCLUSIONS: The conventional in-house developed methods for non-invasive sample collection and DNA extraction from these samples are effective and negate the use of more expensive commercial kits. In conclusion, DNA extracted from mouthwash, saliva, and buccal cytobrush samples provided sufficiently high amounts of pure DNA suitable for molecular analysis of β-thalassemia.

Matched MeSH terms: Specimen Handling/instrumentation; Specimen Handling/methods*
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
Paper-based sample-to-answer molecular diagnostic platform for point-of-care diagnostics

Choi JR, Tang R, Wang S, Wan Abas WA, Pingguan-Murphy B, Xu F

Biosens Bioelectron, 2015 Dec 15;74:427-39.
PMID: 26164488 DOI: 10.1016/j.bios.2015.06.065

Nucleic acid testing (NAT), as a molecular diagnostic technique, including nucleic acid extraction, amplification and detection, plays a fundamental role in medical diagnosis for timely medical treatment. However, current NAT technologies require relatively high-end instrumentation, skilled personnel, and are time-consuming. These drawbacks mean conventional NAT becomes impractical in many resource-limited disease-endemic settings, leading to an urgent need to develop a fast and portable NAT diagnostic tool. Paper-based devices are typically robust, cost-effective and user-friendly, holding a great potential for NAT at the point of care. In view of the escalating demand for the low cost diagnostic devices, we highlight the beneficial use of paper as a platform for NAT, the current state of its development, and the existing challenges preventing its widespread use. We suggest a strategy involving integrating all three steps of NAT into one single paper-based sample-to-answer diagnostic device for rapid medical diagnostics in the near future.

Matched MeSH terms: Specimen Handling/instrumentation
Fulltext Parasitic Gregarine blattarum Found Infecting American Cockroaches, Periplaneta americana, in a Population in Pulau Pinang, Malaysia

Yahaya ZS, Izzaudin NA, Razak AF

Trop Life Sci Res, 2017 Jan;28(1):145-149.
PMID: 28228922 MyJurnal DOI: 10.21315/tlsr2017.28.1.10

A study on the prevalence of a common endoparasite in the wild population of the American cockroach was conducted in Penang Island using a trapping method at several sampling sites on the island. Gregarine blattarum was found in the digestive tract in 5 out of 115, or 4.35%, of the wild American cockroaches, Periplaneta americana, that were sampled. This is the first report in Malaysia of Gregarine blattarum in local American cockroaches.

Matched MeSH terms: Specimen Handling
Fulltext Performance of a Novel Low-Cost, Instrument-Free Plasma Separation Device for HIV Viral Load Quantification and Determination of Treatment Failure in People Living with HIV in Malaysia: a Diagnostic Accuracy Study

Pham MD, Haile BA, Azwa I, Kamarulzaman A, Raman N, Saeidi A, et al.

J Clin Microbiol, 2019 04;57(4).
PMID: 30700508 DOI: 10.1128/JCM.01683-18

HIV viral load (VL) testing is the recommended method for monitoring the response of people living with HIV and receiving antiretroviral therapy (ART). The availability of standard plasma VL testing in low- and middle-income countries (LMICs), and access to this testing, are limited by the need to use fresh plasma. Good specimen collection methods for HIV VL testing that are applicable to resource-constrained settings are needed. We assessed the diagnostic performance of the filtered dried plasma spot (FDPS), created using the newly developed, instrument-free VLPlasma device, in identifying treatment failure at a VL threshold of 1,000 copies/ml in fresh plasma. Performance was compared with that of the conventional dried blood spot (DBS). Venous blood samples from 201 people living with HIV and attending an infectious disease clinic in Malaysia were collected, and HIV VL was quantified using fresh plasma (the reference standard), FDPS, and DBS specimens. VL testing was done using the Roche Cobas AmpliPrep/Cobas TaqMan v2.0 assay. At a threshold of 1,000 copies/ml, the diagnostic performance of the FDPS was superior (sensitivity, 100% [95% confidence interval {CI}, 89.1 to 100%]; specificity, 100% [95% CI, 97.8 to 100%]) to that of the DBS (sensitivity, 100% [95% CI, 89.4 to 100%]; specificity, 36.8% [95% CI, 29.4 to 44.7%]) (P

Matched MeSH terms: Specimen Handling
Fulltext Preserve a Voucher Specimen! The Critical Need for Integrating Natural History Collections in Infectious Disease Studies

Thompson CW, Phelps KL, Allard MW, Cook JA, Dunnum JL, Ferguson AW, et al.

mBio, 2021 Jan 12;12(1).
PMID: 33436435 DOI: 10.1128/mBio.02698-20

Despite being nearly 10 months into the COVID-19 (coronavirus disease 2019) pandemic, the definitive animal host for SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), the causal agent of COVID-19, remains unknown. Unfortunately, similar problems exist for other betacoronaviruses, and no vouchered specimens exist to corroborate host species identification for most of these pathogens. This most basic information is critical to the full understanding and mitigation of emerging zoonotic diseases. To overcome this hurdle, we recommend that host-pathogen researchers adopt vouchering practices and collaborate with natural history collections to permanently archive microbiological samples and host specimens. Vouchered specimens and associated samples provide both repeatability and extension to host-pathogen studies, and using them mobilizes a large workforce (i.e., biodiversity scientists) to assist in pandemic preparedness. We review several well-known examples that successfully integrate host-pathogen research with natural history collections (e.g., yellow fever, hantaviruses, helminths). However, vouchering remains an underutilized practice in such studies. Using an online survey, we assessed vouchering practices used by microbiologists (e.g., bacteriologists, parasitologists, virologists) in host-pathogen research. A much greater number of respondents permanently archive microbiological samples than archive host specimens, and less than half of respondents voucher host specimens from which microbiological samples were lethally collected. To foster collaborations between microbiologists and natural history collections, we provide recommendations for integrating vouchering techniques and archiving of microbiological samples into host-pathogen studies. This integrative approach exemplifies the premise underlying One Health initiatives, providing critical infrastructure for addressing related issues ranging from public health to global climate change and the biodiversity crisis.

Matched MeSH terms: Specimen Handling
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
Quantity and quality assessment of DNA extracted from saliva and blood

Looi ML, Zakaria H, Osman J, Jamal R

Clin. Lab., 2012;58(3-4):307-12.
PMID: 22582505

Saliva has been suggested as an attractive resource for evaluating physiological and pathological conditions in humans. This study aims to evaluate saliva sampling as an alternative to blood sampling for molecular testing.

Matched MeSH terms: Specimen Handling/methods*
Recovery of forensically important entomological specimens from human cadavers in Malaysia--an update

Lee HL

Malays J Pathol, 1989 Aug;11:33-6.
PMID: 2632998

A total of 101 entomological specimens recovered from human cadavers were processed and studied. Analysis of the data indicated that about 95% of these specimens were maggots of flies. Maggots of the blowfly Chrysomya (Family: Calliphoridae) especially Ch. rufifacis and Ch. megacephala were predominantly found in 77 cases (76.2%) while larvae of several other flies of the genera Sarcophaga, Calliphora, Lucilia and hermetia were also recovered. It was notable that Musca domestica or other related flies were not found in all these specimens. The age of these larvae was useful in the determination of the minimum time lapsed after death. However, more biological studies on animal carcases should be conducted for more accurate determinations. Methods of collection, preservation and despatching of specimens were also discussed.

Matched MeSH terms: Specimen Handling*

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