• Title/Summary/Keyword: immunoassay

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Changes of Serum antibody Levels by Indirect Hemagglutination Test after Treatment in Clonorchiasis (IHA(Indirect hemagglutination test)를 이용한 간흡충증(肝吸虫症)의 면역혈청학적(免疫血淸學的) 진단(診斷)에 관한 연구(硏究) -ELISA법과의 비교(比較) 및 치료(治療) 전후(前後) 의 혈청항체가(血淸抗體價) 변동(變動)에 관하여-)

  • Joo, Kyoung-Hwan;Kim, Jong-Seong;Chung, Myung-Sook;Rim, Han-Jong
    • Journal of agricultural medicine and community health
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    • v.12 no.1
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    • pp.117-123
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    • 1987
  • Detection of IgG antibody in clonorchiasis has been accomplished through various serodiagnostic procedure including complement fixation test, gel diffusion test, indirect fluorescent antibody test, indirect hemagglutination test etc. In this report enzyme immunoassay (ELISA) and indirect hemagglutination test (IHA) were used to determine IgG serum antibody levels before and after therapy with praziquantel. Briefly, sera from 62 cases of confirmed human clonorchiasis were examined before and after treatment with praziquantel. Among 62 cases treated 25 cases were categorized as completely cured groups by formalin-ether and careful examination of 4 cellophane thick smered slides at 18 months after treatment. The sera of 25 cases of cured groups were examined again by ELISA and IHA, and com-pared to the previous data. The results obtained were as follows; 1) Sensitivity of IHA test was 83.6% when cut-off titer of 1:8 was applied. No sera obtained from 10 normal healthy control showed positive reaction. 2) Twenty cases (80.0%) out of 25 cured one showed negative results by IHA at 18 months after treatment. 3) Although 5 cases showed positive titer even 18 months after treatment 3 cases of them showed decreased antibody titer. However 2 cases did not show any response. 4) Even though almost all cases showed de- creased ELISA value, only 11 cases (44.0%) out of 25 patients showed negative results by ELISA at 18 months after treatment. In conclusion, it is suggested that, while IgG ELISA for detecting long persisting antibody was more sensitive than IHA, IHA results more conclusively indicated effective treatment in clonorchiasis by negative conversion than did the results of ELISA.

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Bioactive characteristics of an implant surface coated with a pH buffering agent: an in vitro study

  • Pae, Hyung-Chul;Kim, Su-Kyoung;Park, Jin-Young;Song, Young Woo;Cha, Jae-Kook;Paik, Jeong-Won;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.49 no.6
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    • pp.366-381
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    • 2019
  • Purpose: The purpose of this study was to evaluate the effectiveness of conventional sandblasted, large-grit, acid-etched (SLA) surface coated with a pH buffering solution based on surface wettability, blood protein adhesion, osteoblast affinity, and platelet adhesion and activation. Methods: Titanium discs and implants with conventional SLA surface (SA), SLA surface in an aqueous calcium chloride solution (CA), and SLA surface with a pH buffering agent (SOI) were prepared. The wetting velocity was measured by the number of threads wetted by blood over an interval of time. Serum albumin adsorption was tested using the bicinchoninic acid assay and by measuring fluorescence intensity. Osteoblast activity assays (osteoblast adhesion, proliferation, differentiation, mineralization, and migration) were also performed, and platelet adhesion and activation assays were conducted. Results: In both the wetting velocity test and the serum albumin adsorption assay, the SOI surface displayed a significantly higher wetting velocity than the SA surface (P=0.000 and P=0.000, respectively). In the osteoblast adhesion, proliferation, differentiation, and mineralization tests, the mean values for SOI were all higher than those for SA and CA. On the osteoblast migration, platelet adhesion, and activation tests, SOI also showed significantly higher values than SA (P=0.040, P=0.000, and P=0.000, respectively). Conclusions: SOI exhibited higher hydrophilicity and affinity for proteins, cells, and platelets than SA. Within the limits of this study, it may be concluded that coating an implant with a pH buffering agent can induce the attachment of platelets, proteins, and cells to the implant surface. Further studies should be conducted to directly compare SOI with other conventional surfaces with regard to its safety and effectiveness in clinical settings.

Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.1-10
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    • 1999
  • This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

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Analysis of Protease and Antiprotease Concentrations in Retired Workers Exposed to Inorganic Dusts

  • Shin, Jae-Hoon;Hwang, Joo-Hwan;Lee, Kyung-Myung;Lee, Jong-Seong;Lee, Jeong-Oh;Choi, Byung-Soon;Kim, In-Sik
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.309-317
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    • 2009
  • Occupational exposure to inorganic dusts such as coal and silica has been identified as a chronic obstructive pulmonary disease (COPD) risk factor. This risk factor causes lung inflammation and protease-antiprotease imbalance. This abnormal inflammatory response of the lung induces parenchymal tissue destruction and leads to progressive airflow limitation that is characteristics of COPD. The aim of this study was to determine the relationship of proteases such as neutrophil elastase (NE) and matrix metalloproteinase (MMP)-9 and antiproteases such as alpha-1 antitrypsin (AAT) and tissue inhibitors of metalloproteinase (TIMP)-1 with lung function. The study population contained 223 retired workers exposed to inorganic dusts. We performed lung function test, including percent of forced expiratory volume in one second ($%FEV_1$) predicted and $%FEV_1$/forced vital capacity (FVC). We analyzed serum MMP-9, AAT, TIMP-1 and plasma NE concentrations by sandwich enzyme immunoassay. NE, AAT, and TIMP-1 concentrations in workers, who had $%FEV_1$<80% predicted, were higher than those of workers who had $%FEV_1{\geq}80%$ (P<0.05). Both AAT and TIMP-1 concentrations in workers with airflow limitation were higher than those of workers with normal airflow (P<0.05). $%FEV_1$ predicted showed significant negative correlation with AAT (r=-0.255, P<0.0l), TIMP-1 (r=-0.232, P<0.01), and NE (r=-0.196, P<0.01). $%FEV_1$/FVC predicted showed significant negative correlation with NE (r=-0.172, P<0.05). From the results of stepwise multiple regression analysis about $%FEV_1$ and $%FEV_1$/FVC, significant independents were NE (r=-0.135, P=0.001) and AAT (r=-0.100, P=0.013) in $%FEV_1$, and NE (r=-0.160, P=0.014) in $%FEV_1$/FVC. In the present study, there were significant correlations between airflow limitation and protease concentration and between airflow limitation and antiprotease concentration. Serum protease and antiprotease concentrations, however, may be affected by the biological and inflammatory responses. It is necessary to evaluate specimens more reflected the effects of proteases and antiproteases in the lung such as lung tissue, bronchoalveolar lavage fluid, and exhaled breath condensate (EBC).

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Comparison of Enzyme-linked Immunosorbent Assay with Reverse Transcription-polymerase Chain Reaction for Identification of Rotavirus in Neonates (신생아 로타바이러스 감염 진단에서 효소면역법과 중합효소연쇄반응과의 비교)

  • Kim, Sung Eun;Kim, Mi Ok;Park, Sun Young;Jung, Won Jo;Ma, Sang Hyeok;Kim, Yun Jung;Kim, Sun Ju
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.113-119
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    • 2000
  • Purpose : Rotavirus is a most common etiologic agent of pediatric gastroenteritis. The standard method to diagnose rotavirus infection was the detection of viral particles in specimens through electron microscopy. But it was complex. Enzyme immunoassay and latex agglutinin are preferred because they are relatively handy, inexpensive and take a short time, in comparison with electron microscopy. However, several reports have shown that the use of ELISA to diagnose rotavirus infection in neonates can result in false positive reactions. The main purpose of this study is to compare ELISA and RT-PCR in the diagnosis of neonatal rotavirus infection. Methods : Data presented in this study were obtained form 123 newborn babies in the nursery of the Fatima Hospital, Masan, Korea, form Jury to December, 1997. We obtained two samples of stool from each of the newborn babies and then performed the Rotazyme test and the RT-PCR. In the Rotazyme test, the results were interpreted according to visual findings. The samples were used for the RT-PCR test after at stock $-30^{\circ}C$ to identify rotavirus group A. The result of the two tests were compared. Results : The informations are divided into 73 males and females. Out of the total informations 15 were transferred from other hospitals. Their average gestational age was $38.5{\pm}1.6$ weeks. The average birth weight was $3134.8{\pm}539gm$. In the Rotazyme test, 75 samples turned out to be positive. Out of them, 55 samples(75.3%) were positive and 18 samples(24.7%) were negative in the RT-PCR. On the other hand, in the Rotazyme test, 50 samples turned out be negative. Out of them, 27 samples(54%) were positive and 23 samples(46%) were negative in the RT-PCR. Conclusion : Rotavirus infection in uncommon in neonates. The diagnosis based on visual findings using Rotazyme test has a disadvantage in the sense that it can result in false positive reactions and false negative reactions in the diagnosis of neonatal rotavirus infection.

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The Anti-inflammatory Effect and DPPH Free Radical Scavenging Capability of Rhizoma drynariae Aqueous Extract (골쇄보 열수추출물 약침액의 항염증반응 및 DPPH 라디칼 소거능 활성에 대한 연구)

  • Kim, Yong-Min;Kim, Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.32 no.4
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    • pp.169-176
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    • 2015
  • Objectives : This study is to investigate the effects of Rhizoma drynariae aqueous extract(RDA) on cell cytotoxicity, Nitric Oxide (NO) and Prostaglandin $E_2(PGE_2)$ production and 1,1-diphenyl-2-picryl ghdrazyl(DPPH) free radical scavenging capability. Methods : Cell cytotoxicity was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The production of NO was measured by Griess assay. The production of $PGE_2$ was measured by immunoassay. And, the anti-oxidant activity was measured by the DPPH method. Results : Cell cytotoxicity in 50, 100, 200 and $400{\mu}g/ml$ RDA did not increase significantly compared to the RDA untreated group. RDA($200{\mu}g/ml$ and $400{\mu}g/ml$) inhibited NO and $PGE_2$ production in lipopolysaccharide-stimulated RAW 264.7 cells. RDA had high DPPH free radical scavenging capability. Conclusions : This study indicates that RDA inhibits NO and $PGE_2$ production in lipopolysaccharide-stimulated RAW 264.7 cells and improve DPPH free radical scavenging capability. RDA may have an anti-inflammation effect and an anti-oxidant activity.

Comparison Study of AAS and ISE Method in the Lithium Analysis of Serum and Urine (혈액 및 소변의 Lithium치 측정에 있어서 AAS법과 ISE법의 비교)

  • Lee, Soo-In;Lee, Chae-Hoon;Kim, Kyung-Dong;Kim, Chung-Sook
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.409-416
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    • 1993
  • In the method for lithium (Li) analysis, flame emission photometry and atomic absorption spectrophotometry (AAS) have been used most frequently. In addition, lithium can be analyzed by ion-selective electrode (ISE) or fluorscence polarization immunoassay. We evaluated the comparison between AAS method based on the principle of absorption of light at 670.8 nm by Li and ISE method based on the principle of voltage difference generated by Li in contact with lithium ionophore. We compared with those obtained by AAS (AA/AE Spectrophotometer 551, Instrumentation Laboratory Co.) and ISE(CSYNCHRON EL-ISE, Beckman Co.) in the serum and urine of 6 patients and evaluated time-related changes of serum lithium concentration after dosing in both methods. The results are summarized as follows : 1. In within-run precision study for lithium concentration, coefficient variations (CVs, %) ranged from 1.34 to 2.17 for AAS and from 0.34 to 0.85 for ISE method. In between-run precision study for lithium concentration, CVs ranged from 1.23 to 1.72 for AAS and from 0.61 to 1.38 for ISE method. 2. The correlation study between AAS and ISE method resulted in Y=0.946X+0.137 (N=32, r=0.933, X=AAS, Y=ISE) for serum lithium and Y=1.092X+0.977 (N=28, r=0.943, X=AAS, Y=ISE) for urine lithium. 3. Time-related changes of serum lithium concentration in both AAS and ISE method resulted in peak serum levels about 2 hours after dosing and then rapidly decreased after the peak serum level and finally arrived at nearly initial levels about 9 hours after dosing. 4. The reference range of serum lithium was found as undetectable level for both AAS and ISE method and the reference range of urine lithium to the urine creatinine was 0-0.00014 mmol/mg(mean 0.00002 mmol/mg) for AAS method.

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Detection and Type-Distribution of Human Papillomavirus in Vulva and Vaginal Abnormal Cytology Lesions and Cancer Tissues from Thai Women

  • Ngamkham, Jarunya;Boonmark, Krittika;Phansri, Thainsang
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1129-1134
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    • 2016
  • Vulva and Vaginal cancers are rare among all gynecological cancers worldwide, including Thailand, and typically affect women in later life. Persistent high risk human papillomavirus (HR-HPV) infection is one of several important causes of cancer development. In this study, we focused on HPV investigation and specific type distribution from Thai women with abnormality lesions and cancers of the vulva and Vaginal. A total of ninety paraffin-embedded samples of vulva and Vaginal abnormalities and cancer cells with histologically confirmed were collected from Thai women, who were diagnosed in 2003-2012 at the National Cancer Institute, Thailand. HPV DNA was detected and genotyped using polymerase chain reaction and enzyme immunoassay with GP5+/bio 6+ consensus specific primers and digoxigenin-labeled specific oligoprobes, respectively. The human ${\beta}$-globin gene was used as an internal control. Overall results represented that HPV frequency was 16/34 (47.1%) and 8/20 (40.0%) samples of vulva with cancer and abnormal cytology lesions, respectively, while, 3/5 (60%) and 16/33 (51.61%) samples of Vaginal cancer and abnormal cytology lesions, respectively, were HPV DNA positive. Single HPV type and multiple HPV type infection could be observed in both type of cancers and abnormal lesion samples in the different histological categorizes. HPV16 was the most frequent type in all cancers and abnormal cytology lesions, whereas HPV 18 was less frequent and could be detected as co-infection with other high risk HPV types. In addition, low risk types such as HPV 6, 11 and 70 could be detected in Vulva cancer and abnormal cytology lesion samples, whereas, all Vaginal cancer samples exhibited only high risk HPV types; HPV 16 and 31. In conclusion, from our results in this study we suggest that women with persistent high risk HPV type infection are at risk of developing vulva and Vaginal cancers and HPV 16 was observed at the highest frequent both of these, similar to the cervical cancer cases. Although the number of samples in this study was limited and might not represent the overall incidence and prevalence in Thai women, but the baseline data are of interest and suggest further study for primary cancer screening and/or developing the efficiency of prophylactic HPV vaccines in Thailand.

Human Epididymis Protein 4 Reference Intervals in a Multiethnic Asian Women Population

  • Mokhtar, N.M.;Thevarajah, M.;M.A., Noorazmi;M., Isahak
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.12
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    • pp.6391-6395
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    • 2012
  • Background: Ovarian cancer is ranked as the fifth most common cause of cancer death in women. In Malaysia, it is the fourth most common cancer in females. CA125 has been the tumor marker of choice in ovarian cancer but its diagnostic specificity in early stages is only 50%. Hence, there is a critical need to identify an alternative tumor marker that is capable of detecting detect ovarian cancer at an early stage. HE4 is a new tumor marker proposed for the early diagnosis of ovarian cancer and disease recurrence. Currently, none of the normal ranges of HE4 quoted in the literature are based on data for a multiethnic Asian population. Therefore, the aim of this study was to determine reference intervals for HE4 in an Asian population presenting in University Malaya Medical Centre, a tertiary reference hospital. Materials and Methods: 300 healthy women were recruited comprising 150 premenopausal and 150 postmenopausal women, aged from 20-76 years. All women were subjected to a pelvic ultrasonograph and were confirmed to be free from ovarian pathology on recruitment. Serum HE4 levels were determined by chemiluminescent microparticle immunoassay (CMIA, Abbott Architect). The reference intervals were determined following CLSI guidelines (C28-A2) using a non-parametric method. Results: The upper limits of the $95^{th}$ percentile reference interval (90%CI) for all the women collectively were 64.6 pmol/L, and 58.4 pmol/L for premenopausal) and 69.0 pmol/L for postmenopausal. The concentration of HE4 was noted to increase with age especially in women who were more than 50 years old. We also noted that our proposed reference limit was lower compared to the level given by manufacturer Abbott Architect HE4 kit insert (58.4 vs 70 pmol/L for premenopausal group and 69.0 vs 140 pmol/L in the postmenopausal group). The study also showed a significant difference in HE4 concentrations between ethnic groups (Malays and Indians). The levels of HE4 in Indians appeared higher than in Malays (p<0.05), while no significant differences were noted between the Malays and Chinese ethnic groups. Conclusions: More data are needed to establish a reference interval that will better represent the multiethnic Malaysian population. Probably a larger sampling size of equal representation of the Malay, Chinese, Indians as well as the other native ethnic communities will give us a greater confidence on whether genetics plays a role in reference interval determination.

Association between Circulating Vitamin D, the Taq1 Vitamin D Receptor Gene Polymorphism and Colorectal Cancer Risk among Jordanians

  • Atoum, Manar Fayiz;Tchoporyan, Melya Nizar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7337-7341
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    • 2014
  • Background: The physiological role of vitamin D extends beyond bone health and calcium-phosphate homeostasis to effects on cancer risk, mainly for colorectal cancer. Vitamin D may have an anticancer effect in colorectal cancer mediated by binding of the active form $1,25(OH)_2D$ to the vitamin D receptor (VDR). The Taq1 VDR gene polymorphism, a C-to-T base substitution (rs731236) in exon 9 may influence its expression and function. The aim of this study wass to determine the 25(OH)D vitamin D level and to investigate the association between circulating vitamin D level and Taq1VDR gene polymorphism among Jordanian colorectal cancer patients. Materials and Methods: This case control study enrolled ninety-three patients and one hundred and two healthy Jordanian volunteers from AL-Basheer Hospital/Amman (2012-2013). Ethical approval and signed consent forms were obtained from all participants before sample collection. 25(OH)D levels were determined by competitive immunoassay Elecsys (Roche Diagnostic, France). DNA was extracted (Promega, USA) and amplified by PCR followed by VDR Taq1 restriction enzyme digestion. The genotype distribution was evaluated by paired t-test and chi-square. Comparison between vitamin D levels among CRC and control were assessed by odds ratio with 95% confidence interval. Results: The vitamin D serum level was significantly lower among colorectal cancer patients (8.34 ng/ml) compared to the healthy control group (21.02ng/ml). Patients deficient in vitamin D (less than 10.0 ng/ml) had increased colorectal cancer risk 19.2 fold compared to control. Only 2.2% of CRC patients had optimal vitamin D compared to 23.5% among healthy control. TT, Tt and tt Taq1 genotype frequencies among CRC cases was 35.5%, 50.5% and 14% compared to 43.1%, 41.2% and 15.7% among healthy control; respectively. CRC patients had lower mean vitamin D level among TT ($8.91{\pm}4.31$) and Tt ($9.15{\pm}5.25$) genotypes compared to control ($21.3{\pm}8.31$) and ($19.3{\pm}7.68$); respectively. Conclusions: There is significant association between low 25(OH)D serum level and colorectal cancer risk. The VDRTaq1 polymorphism was associated with increased colorectal cancer risk among patient with VDRTaq1 TT and Tt genotypes. Understanding the functional mechanism of VDRTaq1 TT and Tt may provide a strategy for colorectal cancer prevention and treatment.