Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay)

Nandhakumar, S.; Parasuraman, Subramani; Shanmugam, MM; Rao, K Ramachandra; Chand, Parkash; Bhat, B. Vishnu

doi:10.4103/0976-500x.81903

Cited by 170 publications

(106 citation statements)

References 13 publications

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“…DISCUSSION the cases. Comet assay variables were resembling normal male as compared with others with primary amenorrhoea [9,10]. Clinical features were similar in both the individuals except for renal agenesis in case 1.…”

Section: Case Descriptionmentioning

confidence: 55%

Androgen Insensitivity Syndrome Among Cousin Sisters- A Rare Entity

Ramamurthy¹,

Karuppusamy²

2018

IJAR

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Androgen insensitivity syndrome (AIS), is a X-linked disorder characterized by resistance to androgen caused by mutation of androgen receptor gene in which XY karyotype individuals exhibit female phenotype.AIS is characterised by evidence of feminization (under masculinization) of the external genitalia at birth, abnormal secondary sexual development at puberty, and infertility in individuals with 46 XY karyotype.We are presenting here a familial case of complete androgen insensitivity syndrome in south Indian Population. 46 XY karyotype was found in two subjects who were cousin sisters with female phenotype,who presented with primary amenorrhoea. Comet assay was done, which showed results comparable with normal males. In both girls' inguinal gonads was present which was removed and hormonal therapy with estrogen was given to prevent osteoporosis. Androgen insensitivity syndrome can be inherited as an X linked disorder as evidenced by previous studies.

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Section: Case Descriptionmentioning

confidence: 55%

Androgen Insensitivity Syndrome Among Cousin Sisters- A Rare Entity

Ramamurthy¹,

Karuppusamy²

2018

IJAR

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“…20) Blood samples (~0.5 mL/participant) were taken in prelabelled microcentrifuge tubes containing heparin sodium (3U, Biological E Limited, India) and processed within 3-4 hours for DNA damage assessment. SCGE/Comet assay: Prior to the SCGE assay, the blood samples were assessed for cell viability by the trypan blue dye exclusion method 21) and since cell viability was 92-100%, the alkaline version of SCGE assay was performed on PBL according to the method of Singh, et al 22) with slight modifications which included use of local chemicals, agarose-precoated slides in lieu of frosted slides, 23) and silver-staining of comets instead of using ethidium bromide. 24) In brief, 110 μL of molten low melting point agarose (LMPA; 0.5%, SRL, India) mixed with 20 μL of whole blood was poured over slides pre-coated with 1% normal melting point agarose (NMPA, SRL, India) and immediately covered with cover slips while avoiding air bubbles and kept at 4°C for 5 minutes to allow the gels to solidify.…”

Section: Methodsmentioning

confidence: 99%

Assessment of DNA Damage in Leukocytes of Patients With Coronary Artery Disease by Comet Assay

Bhat

Gandhi

2017

Int. Heart J.

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SummaryDNA damage in the peripheral blood leukocytes (PBL) of patients with coronary artery disease (CAD) was investigated using the sensitive alkaline single cell gel electrophoresis (SCGE)/comet assay.This case-control study consisted of CAD patients (n = 200; mean age, 59.04 ± 0.75 years) undergoing treatment at local hospitals and age-, sex-, and ethnicity-matched healthy controls (n = 200; mean age, 57.88 ± 0.96 years) from the general population.CAD patients had significantly (P < 0.001) increased DNA damage (tail DNA percent (T-DNA %) 22.45 ± 0.50 versus 5.81 ± 0.28; tail moment (TM) 89.35 ± 3.16 versus 9.98 ± 0.69; Olive tail moment (OTM) 60.50 ± 1.79 versus 10.94 ± 0.63; damage frequency (DF) 91.12 ± 0.93 versus 41.78 ± 2.04, damage index (DI) 173.68 ± 3.36 versus 48.53 ± 2.59) compared to controls. Patients with acute myocardial infarction (AMI) showed significantly higher DNA damage than patients with unstable angina (UA) (T-DNA % 24.05 ± 0.87 versus 21.06 ± 0.90; TM 100.02 ± 6.19 versus 81.61 ± 5.84; OTM 66.19 ± 3.20 versus 56.47 ± 3.33; DF 94.02 ± 0.84 versus 91.10 ± 1.16, DI 184.13 ± 5.33 versus 166.42 ± 5.89). Moreover, DNA damage was found to be significantly (P < 0.05) elevated in patients receiving ecosprin, ramipril, and metoprolol therapy compared to aspirin and nitrocontin.The increased DNA damage in CAD patients may be the consequence of disease and/or drug therapy. These observations are of concern because unrepaired DNA can lead to malignancy, and the likelihood of increasing mortality and morbidity rates in CAD patients. (Int Heart J 2017; 58: 271-274)

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“…Comet assay, a simple, rapid, visual and biochemical technique, was used for the detection of various genotoxic damages in mammalian cells [20]. It has the ability to detect single/double-strand DNA breaks, DNA crosslink, oxidation and alkylation of bases, base/base-pair damages and apoptotic nuclei in eukaryotic cells [19]. DNA damage can result in multiple lesions including mutations, insertions, deletions, translocations, and loss of chromosomes and essential genetic information.…”

Section: Tablementioning

confidence: 99%

“…The Komet 5 image analysis software (Kinetic Imaging, Ltd. Liverpool, UK) linked to a charge-coupled device camera was used to determine the quantitative and qualitative extent of DNA damage in the liver cells by measuring the length of DNA migration, migrated DNA percentage and tail moment through the observation of fifty to hundred cells per sample [19]. Tailed cells indicated by the ratio of the number of comet tails and the number of non-head shapes to the number of total cells.…”

Section: Assessment Of Hepatocyte Cell's Genotoxicity By Comet Assaymentioning

confidence: 99%

Hepatoprotective activity of Thymus vulgaris extract against Toxoplasma gondii infection

El-Sayed¹

2017

APJTD

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Evaluation of DNA damage using single-cell gel electrophoresis (Comet Assay)

Cited by 170 publications

References 13 publications

Androgen Insensitivity Syndrome Among Cousin Sisters- A Rare Entity

Androgen Insensitivity Syndrome Among Cousin Sisters- A Rare Entity

Assessment of DNA Damage in Leukocytes of Patients With Coronary Artery Disease by Comet Assay

Hepatoprotective activity of Thymus vulgaris extract against Toxoplasma gondii infection

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