American Journal of Microbiological Research

ISSN (Print): 2328-4129

ISSN (Online): 2328-4137



Molecular Characterization and in Silico Analysis of a Novel Mutation in TEM-1 Beta-Lactamase Gene among Pathogenic E. coli infecting a Sudanese Patient

1Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan

2Department of microbiology, Faculty of medicine, university of Khartoum, Sudan

3Head department of biotechnology, Biotechnology Park, Africa city of technology, Sudan

4Botany department, Faculty of Science, University of Khartoum, Sudan

American Journal of Microbiological Research. 2014, 2(6), 217-223
DOI: 10.12691/ajmr-2-6-8
Copyright © 2014 Science and Education Publishing

Cite this paper:
Hisham N Altayb, Nagwa M El Amin, Maowia M. Mukhtar, Mohamed Ahmed Salih, Mohamed A M Siddig. Molecular Characterization and in Silico Analysis of a Novel Mutation in TEM-1 Beta-Lactamase Gene among Pathogenic E. coli infecting a Sudanese Patient. American Journal of Microbiological Research. 2014; 2(6):217-223. doi: 10.12691/ajmr-2-6-8.

Correspondence to: Hisham  N Altayb, Institute of Endemic Diseases, University of Khartoum, Khartoum, Sudan. Email:


The presence of ESBLs in many E. coli strains are of serious concern, since these organisms are the most common cause of different human infections. In this study we isolate an E. coli bacterium with high hydrolytic activity against cefotaxime. The ESBLs production was confirmed by phenotypic confirmatory test, while the ESBLs genes were detected by polymerase chain reaction (PCR). This isolate was positive for TEM gene and negative for CTX-M and SHV genes. DNA sequencing was done for TEM gene. The nucleotide sequences and translated proteins were subjected to BLAST for sequences similarity and homology, BLASTp result revealed a substitution of aspartic acid in TEM-1(gb: AFI61435.1) to Threonine at position 262. In Silico tools was used for mutation analysis and prediction of secondary and tertiary structure of wild and mutant type genes. We conclude that our mutant gene is completely different from the wild types TEM-1 gene, within phenotypic and genotypic levels. So we conclude a novel mutant TEM gene with ESBLs activity is been detected in Sudan Phylogenetic tree revealed that the possible source of our gene is Iran.



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Detection of Cryptosporidium oocyts in Commonly Consumed Fresh Salad Vegetables

1Department of Food Processing and Preservation, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

2Department of Agricultural chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

3Department of Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

American Journal of Microbiological Research. 2014, 2(6), 224-226
DOI: 10.12691/ajmr-2-6-9
Copyright © 2014 Science and Education Publishing

Cite this paper:
Jiaur Rahman, Aminul Islam Talukder, Farid Hossain, Sultan Mahomud, M. Atikul Islam, Shamsuzzoha. Detection of Cryptosporidium oocyts in Commonly Consumed Fresh Salad Vegetables. American Journal of Microbiological Research. 2014; 2(6):224-226. doi: 10.12691/ajmr-2-6-9.

Correspondence to: Jiaur  Rahman, Department of Food Processing and Preservation, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh. Email:


The aim of this study was to determine the degree of contamination caused by Cryptosporidium oocyts in regularly consumed salad vegetables sold at various wholesale and retail markets in northern part of Bangladesh. A total number of 165 samples of salad vegetables collected from different wholesale and retail markets were examined for detection of Cryptosporidium oocysts using sucrose flotation medium of 1.18 specific gravity and Ziehl Neelsen staining technique with some modifications. Cryptosporidium oocysts were detected in 47 (30%) of the total examined samples. About 40 Tomato, 35 Cucumber, 20 Lettuce, 35 Carrot and 35 Mint’s leaf samples were examined while Lettuce had the highest (40%) contamination rate followed by Tomato (32.5%), Carrot (31.4%), Cucumber (25.7%), and Mint’s leaf (22.8%). There was no significant difference (x2 = 2.278; p <0.05) among occurrences of Cryptosporidium oocysts in usually consumed salad vegetables sold at market. This study has shown that salad vegetables sold at wholesale and retail markets in northern part of Bangladesh are contaminated with Cryptosporidium oocysts, may pose a health risk to consumers of such products. This reveals food safety and significance of public health.



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Molecular Characteristic and Insilico Analysis of KatG Gene in Isoniazid Resistance Mycobacterium Tuberculosis Isolate from Sudan

1Department of Microbiology, university of Bahri, Sudan

2Department of Microbiology, Khartoum College of Medical Sciences, Sudan

3Department of Bioinformatics, Africa City of Technology, Sudan

4Department of Computer Sciences, Najran University, KSA

5Department of Histopathology, ALzaim AL-Azhari University, Sudan

6Department of Microbiology Al borg Medical Laboratories, UAE

7Department of environmental Health, University of Hail, KSA

8Division of Molecular Genetics, University of Tuebingen, Germany;University, HNO –universities Klink-Tuebingen (Germany)

American Journal of Microbiological Research. 2014, 2(6), 227-233
DOI: 10.12691/ajmr-2-6-10
Copyright © 2014 Science and Education Publishing

Cite this paper:
Gusai H.Abdel Samad, Solima M. A. Sabeel, Walaa A. Abuelgassim, Abeer E. Abdelltif, Wisam M. Osman, Mona A. Haroun, Somaya M. Soliman, Sami. A. B. Salam, Hamid. A. Hamdan, Mohamed A. Hassan. Molecular Characteristic and Insilico Analysis of KatG Gene in Isoniazid Resistance Mycobacterium Tuberculosis Isolate from Sudan. American Journal of Microbiological Research. 2014; 2(6):227-233. doi: 10.12691/ajmr-2-6-10.

Correspondence to: Solima  M. A. Sabeel, Department of Microbiology, Khartoum College of Medical Sciences, Sudan. Email:


The KatG gene of Mycobacterium Tuberculosis has been associated with isoniazid (INH) drug resistance. While isoniazid (INH) considered as corner stone and main chemotherapy used throughout the world to manage tuberculosis, thus the Progress in apprehension of principle concepts associated with resistance to isoniazid (INH) has allowed molecular tests in addition to bioinformatics tool for the detection of drug-resistant tuberculosis to be developed. In Consecutive isolates (n = 20) of multidrug-resistant Mycobacterium tuberculosis, part of the katG was sequenced for INH resistance analysis. BLAST analysis of all sequences revealed 100% identity with the available strain “EGY-K361” Mycobacterium tuberculosis with Accession No: KC49137.1 except 6 isolates :isolate1, 2, 4, 11, 15, and isolate 20 revealed 99% identity. Thosesix isolates (30%) have detected mutation in Catalase-peroxidase enzyme S315T; three isolate from six 3/6 (50%) of mutant isolates have SNP AGC>ACC substitution while others 3/6 have substitution C>G in position 1280 which may contributed in altering gene expression. The secondary structure of wild and mutant proteins had been done using phyre2 software while the three dimensional structures of them had been done by Chimera software. Stability of mutant protein was increased which detected by i-mutant. Phylogenetic tree of the sequences revealed two distinct phylogroups: mutant isolates and wild isolates phylogroups with controls from different countries retrieved from Gene bank. Serine at position 315 is one of potential drug active sites that proved via SiteEngine soft ware, therefore any substitution will change efficiency of INH.



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Effect of Some Non steroidal Anti-Inflammatory Drugs on Growth, Adherence and Mature Biofilms of Candida spp.

1Undergraduate student, faculty of pharmacy, Minia university

2Undergraduate student, Faculty of science, Minia university

3Demonstrator, Department of Microbiology, Faculty of Pharmacy, Minia university

4A researcher, Department of Microbiology, Faculty of Pharmacy, Minia university

5Lecturer of Microbiology, Department of Microbiology, Faculty of Pharmacy, Minia university

American Journal of Microbiological Research. 2015, 3(1), 1-7
DOI: 10.12691/ajmr-3-1-1
Copyright © 2015 Science and Education Publishing

Cite this paper:
Ahmad Ashraf, Fatma Yousri, Nora Taha, Omar Abd El-Waly, Abd El-Kareem Ramadan, Esraa Ismail, Reham Hamada, Mohamed Khalaf, Mohamed Refaee, Sameh Ali, Abobakr Madyn, Rehab Mahmoud Abd El-Baky. Effect of Some Non steroidal Anti-Inflammatory Drugs on Growth, Adherence and Mature Biofilms of Candida spp.. American Journal of Microbiological Research. 2015; 3(1):1-7. doi: 10.12691/ajmr-3-1-1.

Correspondence to: Rehab  Mahmoud Abd El-Baky, Lecturer of Microbiology, Department of Microbiology, Faculty of Pharmacy, Minia university. Email:


Candida spp. are the most common cause of fungal diseases and the fourth commonest cause of nosocomial invasive infections which are considered in many cases as life threatening. Among Candida spp., C. albicans is the most common cause of many fungal diseases, but non-albicans spp. infections are in increase. Non-steroidal anti-inflammatory drugs have previously been shown to have antifungal activity. In this study we determine the antifungal activity of the tested NSAIDs using agar well diffusion method, their effect on the dimorphic transition of C. albicans by testing their ability to form germ tube in the presence of human serum. Determining the effect of NSAIDs on the adherence to plastic surfaces and on the mature biofilms formed by the tested Candida spp.. The results indicated that Sodium Diclofenac showed lower MIC against C. albicans and C. glabrata while Ibuprofen had lower MIC against C. krusei. upon examining the effect of Diclofenac sodium, Ibuprofen and ketoprofen on biofilm formed on polyurethane segments by Scanning electron microscope (SEM), a damage to membranes of the tested species was observed. Sodium Diclofenac showed the highest inhibitory effect on the adherence (51.1-76.9% at MIC and 56.6-83.3% at 2XMIC) of C. albicans and C. glabrata but Ibuprofen showed a higher inhibitory effect against the adherence of C. krusei. For mature biofilms, the highest disruptive effect on mature biofilms formed by all tested Candida Spp. (37.72-59.29% at MIC and 42.68-63.06% at 2XMIC) was observed by Diclofenac sodium. Sodium Diclofenac inhibited dimorphic transition of C. albicans but a decrease in germ tube formation was shown by others. In conclusion, the tested drugs showed antifungal, anti-adherent and anti-biofilm activity that make them useful in the treatment of fungal infection and the prevention of biofilm formation on the surface of medical devices.



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The Effect of the Petroleum Ether Extracts from Mangosteen Pericarp (Garcinia mangostana L.) on Interferon-gammaand, Interleukin-12 Activities in AlbinoWistar Rats (Rattus norvegicus) Infected with Mycobacterium tuberculosis

1Biology Study Programm, Faculty of Mathematic and Sciences, Pattimura University, Ambon, Indonesia

2Department of Internal Medicine, Faculty of Medicine, Hasanudin University, Makassar, Indonesia

3Department of Microbiology, Faculty of Medicine, Mulawarman University, Samarinda, Indonesia

4Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia

American Journal of Microbiological Research. 2015, 3(1), 8-13
DOI: 10.12691/ajmr-3-1-2
Copyright © 2015 Science and Education Publishing

Cite this paper:
Martha Kaihena, Syamsu, Yadi Yasir, Mochammad Hatta. The Effect of the Petroleum Ether Extracts from Mangosteen Pericarp (Garcinia mangostana L.) on Interferon-gammaand, Interleukin-12 Activities in AlbinoWistar Rats (Rattus norvegicus) Infected with Mycobacterium tuberculosis. American Journal of Microbiological Research. 2015; 3(1):8-13. doi: 10.12691/ajmr-3-1-2.

Correspondence to: Mochammad  Hatta, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Makassar, Indonesia. Email:


Background. Garcinia mangostana L(GML) pericarp extract is known to contain active substances called Xanthones (α, β and γ Mangosteens) which is the biggest derivative and has strong antioxidant effects. This substance also has antiinflammatory, antilipid, anticancer, antibacteria and antituberculosis effects. However, its mechanism is still unclear. Mycobacterium tuberculosis (M. tuberculosis) invasion into the lungs through the respiratory tract can cause severe infection. The body has an immune system which controls infection by eliminating germs to ease the burden of infection. Interferon gamma (IFN- γ) and Interleukin 12 (IL-12) acts as positive feedback in stimulating macrophages to kill M. tuberculosis. During this process, oxidative compounds (ROI,RNI,NO) that plays an important role in the phagolysosome fusion process are produced. Not only does the consumption of GML pericarp extract as an antioxidant becomes the immunomodulator to enhance the immune’s activities, it also functions as an antioxidant that can neutralize the oxidative compounds produced by the immune system. The aim of this study was to determine the effect of EPEBh GML on IFN-γ and IL-12 secretion activities in mice infected with M. tuberculosis. Materials and method. This study used 30 Wistar rats, 150-200g of weight and 8-10 weeks old. Rats were randomly divided into 6 groups each consisting of 5 rats, including the negative control (without infection and without EPEKBh GML intervention) and a positive control (rats were infected with M.tb H37Rv at a dose of 106cfuas much as 0.2 ml through the trachea for 6 weeks). Once infected, the rats were then intervened with EPEKBhGML 30, 60, 120 dose and 180 mg/kg bodyweigh/day for 1 month. Afterward the rats were necropsied and dissected for the blood to be taken directly from the heart. Levels of IFN-γ and IL-12 were analyzed using the ELISA method. Data were then analyzed using One-Way ANOVAtest followed by Post-Hoc test (LSD), a significant P <0.05, to assess the comparison between groups. Results. Results show that EPEKBh GML significantly affect the rise of IFN-γ levels, with a P value 0,000<0,05 and IL-12, with a P value of 0,045 <0,05. Conclusion. EPEKBh GML was effective in increasing the activity of IFN-γ and IL-12 and the most effective dose to increase IFN-γ and IL-12activity was 120 mg/kgbody weight/day. Also, increasing the dose to a higher dosage had no effect on IFN-γ and IL-12 activity, in fact it tended to decline.This results need to further study to understand what the reason.



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Relationship between Temperature, Ph and Population of Selected Microbial Indicators during Anaerobic Digestion of Guinea Grass (Panicum Maximum)

1Department of Microbiology, Faculty of Biological Science, College of Natural and Applied Sciences, University of Port Harcourt, P.M.B. 5323 Port Harcourt, Nigeria

American Journal of Microbiological Research. 2015, 3(1), 14-24
DOI: 10.12691/ajmr-3-1-3
Copyright © 2015 Science and Education Publishing

Cite this paper:
Ogbonna. C. B., Berebon. D. P., Onwuegbu. E. K.. Relationship between Temperature, Ph and Population of Selected Microbial Indicators during Anaerobic Digestion of Guinea Grass (Panicum Maximum). American Journal of Microbiological Research. 2015; 3(1):14-24. doi: 10.12691/ajmr-3-1-3.

Correspondence to: Ogbonna.  C. B., Department of Microbiology, Faculty of Biological Science, College of Natural and Applied Sciences, University of Port Harcourt, P.M.B. 5323 Port Harcourt, Nigeria. Email:


In this study, the relationship between process temperature, process pH and population of selected microbial indicators during anaerobic digestion of guinea grass (Panicum maximum) at ambient condition was investigated. A one stage batch-type mesophilic anaerobic digestion system was configured using rumen fluid (RF) as inoculums (ADRF) and a low solid loading of approximately 7.0% total solid (TS). Physicochemical parameters such as process temperature (PTMRF), process pHRF and volatile fatty acid (VFARF) were monitored with time. Selected indicator microbial populations were monitored by standard cultural techniques based on metabolic capacity and oxygen sensitivity with respect to time. Result showed that average PTMRF increased from 27.5°C to 35.2°C, average process pHRF ranged from 6.5 to 7.9 and VFARF ranged from 1,080.00 mg/L to 4,800.33 mg/L with time. In terms of metabolic capacity and oxygen sensitivity, the populations of cellulolytic bacteria (CBRF), lactose and glucose fermenting (acidogenic) bacteria (LFBRF and GFBRF), propionate and ethanol oxidizing (acetogenic) bacteria (POBRF and EOBRF), acetate oxidizing methanogens (AOMRF), obligate anaerobic bacteria (OABRF) and total facultative bacteria (FAABRF) increased (about 10-fold) respectively with time. Correlation analysis showed positive relationships between the process temperature (PTMRF) and the population of selected microbial indicators with time. However, there were negative relationships between the process pHRF and the population of selected microbial indicators with time. Furthermore, there were positive relationships between the populations of selected microbial indicators with time. Rumen fluid significantly (P < 0.05) affected the dynamics of the process temperature (PTMRF) and process pHRF inside the ADRF system with time respectively. These kinds of relationships between biotic factors and between biotic and abiotic factors could be used to monitor the state of anaerobic digestion process with respect to time.



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Development of an in Vitro Novel Device that Simulates the Real Life of the Biofilm Formation on Catheters under both Static and Continuous Fluid Flow Systems

1Department of Microbiology, Immunology and Biotechnology, Faculty of Pharmacy and Biotechnology, German University in Cairo, GUC, New Cairo City, Egypt

American Journal of Microbiological Research. 2015, 3(1), 25-32
DOI: 10.12691/ajmr-3-1-4
Copyright © 2015 Science and Education Publishing

Cite this paper:
Mohamed El-Azizi. Development of an in Vitro Novel Device that Simulates the Real Life of the Biofilm Formation on Catheters under both Static and Continuous Fluid Flow Systems. American Journal of Microbiological Research. 2015; 3(1):25-32. doi: 10.12691/ajmr-3-1-4.

Correspondence to: Mohamed  El-Azizi, Department of Microbiology, Immunology and Biotechnology, Faculty of Pharmacy and Biotechnology, German University in Cairo, GUC, New Cairo City, Egypt. Email:


Biofilm model systems are essential to explore the development and the nature of the microbial community within the biofilm as well as the mechanism of their resistance. The aim of this work is to develop a simple in vitro novel device which mimics the real life of the biofilm formation and could be modulated to contain most catheter and tubes and readily allows biofilm formation under different experimental conditions. Two clinical isolates, Staphylococcus epidermidis and Candida albicans, were used to validate the device. The viability of the microorganisms within the biofilm was demonstrated quantitatively by viable count and semi-quantitively by using Scanning Electron Microscope and Confocal Scanning Laser Microscope. The shear stress on the inner and outer surfaces of the catheter was determined at different flow rates of the culture medium. The presented device supports biofilm formation of the tested microorganisms under static and dynamic fluid flow systems. The results are comparable to that of other biofilm models. The number of cells contained in the biofilm under static system was significantly higher than that of the biofilm which formed under dynamic system for both microorganisms. For S. epidermidis, the log value of the number of cells contained in the biofilm under static system was 6.41 ± 0.22 compared to 5.18 ±0.13 of the biofilm which formed under continuous fluid flow system (p < 0.001). For C. albicans, the log value of the number of adherent cells was 6.44 ± 38 and 5.47 ± 0.05 respectively (p = 0.012). The presented well suited to study the real life of the biofilm formation by microorganisms. It enables the formation of a reproducible biofilm of bacteria and yeast on the catheter surface in both static and dynamic systems and its design permits low laminar flow system.



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Antibodies to Echinococcus granulosus Egg Antigens in Hydatid Patients: Characterization by Immunoblot and Diagnostic Value

1Laboratory of Immunology, Institut Pasteur de Tunis, Tunis, TUNISIA

American Journal of Microbiological Research. 2015, 3(1), 33-40
DOI: 10.12691/ajmr-3-1-5
Copyright © 2015 Science and Education Publishing

Cite this paper:
Wissal IRAQI HOUSSAINI. Antibodies to Echinococcus granulosus Egg Antigens in Hydatid Patients: Characterization by Immunoblot and Diagnostic Value. American Journal of Microbiological Research. 2015; 3(1):33-40. doi: 10.12691/ajmr-3-1-5.

Correspondence to: Wissal  IRAQI HOUSSAINI, Laboratory of Immunology, Institut Pasteur de Tunis, Tunis, TUNISIA. Email:


Cystic echinococcosis/hydatidosis, caused by Echinococcus granulosus, is a chronic zoonotic larval cestode infection in humans. Human hydatidosis is a public health parasitic disease that is cosmopolitan in its distribution. Several years may last between contamination with E. granulosus eggs and development of symptomatic cysts. Antigens expressed by embryos at early differentiation steps between the embryo and the young cyst may be interesting as markers of early infection or reinfection. This study investigates antibody response (IgG) to E. granulosus egg antigens by Immunoblot (IB) and ELISA in 20 hydatid patients surgically confirmed. We have also tested sera using protoscoleces and hydatid cyst fluid (HCF) from E. granulosus. The results show that hydatid patients reacted to eggs as well as protoscoleces and HCF. However, eggs share antigens in common with other life-cycle stages, but also support the notion that they may possess some unique stage-specific antigens determinants. The possible functional significance and the potential use of combined assessment of these antigens in IB with Ig G response in ELISA are discussed.



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Evaluation of the Degree of Contamination of Salad Vegetables Sold in Umuahia Main Market

1Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture Umudike, Abia State, Nigeria.

American Journal of Microbiological Research. 2015, 3(1), 41-44
DOI: 10.12691/ajmr-3-1-6
Copyright © 2015 Science and Education Publishing

Cite this paper:
Nwankwo. I.U, Eze. V.C, Onwuakor. C.E, Friday. J.U. Evaluation of the Degree of Contamination of Salad Vegetables Sold in Umuahia Main Market. American Journal of Microbiological Research. 2015; 3(1):41-44. doi: 10.12691/ajmr-3-1-6.

Correspondence to: Onwuakor.  C.E, Department of Microbiology, College of Natural Sciences, Michael Okpara University of Agriculture Umudike, Abia State, Nigeria.. Email:


The evaluation of the degree of contamination of salad vegetables sold in Umuahia main market was carried out. Six Samples of salad vegetables (Cabbage, Carrot, Cucumber, Onions, Green peas and Green beans) sold around the market were analyzed. Total viable counts ranged from 2.35 x 105 to 9.50 x 105 cfu/ml with highest incidence seen in cabbage and onions sold by vendor A. 86.66% of cabbage samples were contaminated, with the least incidence seen in green bean (33.33%). Six bacterial and five fungal species were isolated. The isolates include: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus sp, Enterobacter sp and Proteus sp. and the fungal isolates were Aspergillus sp. Yeast, Cladosporium sp. Penicillium sp. and Rhizopus sp. These results indicates that street vended salad vegetables where contaminated to an extent, and therefore, needs vigorous washing with safe running water before consuming to avoid food borne diseases associated with consumption of contaminated food.



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Development of SCAR marker for Specific Detection of Trichoderma harzanium and Trichoderma viride

1Department of Biotechnology, College of Agriculture, Junagadh Agriculture University, Junagadh -362001, India

2Scientific Research Center, Biotechnology and Genetic Engineering Unit, Taif University, KSA

3Genetics Department, Faculty of Agriculture, Minufiya University, Egypt

4Microbiology Dept, MVM Science and Home Science College, Saurashtra University, Rajkot -360001, India

American Journal of Microbiological Research. 2015, 3(1), 45-49
DOI: 10.12691/ajmr-3-1-7
Copyright © 2015 Science and Education Publishing

Cite this paper:
H. J. Parmar, M. M. Hassan, N. P. Bodar, H .N. Lakhani, V. V. Umrania, B. A. Golakiya. Development of SCAR marker for Specific Detection of Trichoderma harzanium and Trichoderma viride. American Journal of Microbiological Research. 2015; 3(1):45-49. doi: 10.12691/ajmr-3-1-7.

Correspondence to: M.  M. Hassan, Scientific Research Center, Biotechnology and Genetic Engineering Unit, Taif University, KSA. Email:


Trichoderma have been used as biological control agents against soil borne plant pathological fungi such as S. rolfsii. On this day molecular based techniques has been developed to detect T. harzanium using a fungus-specific marker derived from genomic DNA. An amplified RAPD product of 220 bp and 900 bp obtained in T. harzanium (NBAII Th 1) and T. viride (NBAII Tv 2) isolates, respectively. These two RAPD products were obtained using two random primers OPA-16 for harzanium (NBAII Th 1) isolate and OPC-05 for T. viride (NBAII Tv 2), thin RAPD products were sequenced. Based on sequences, one primers were designed, out of which a primer pair HAR220FP5 (CTTTTGGTTTGACACGGTTCT and HAR220RP5 (AAGCTTTGAAGTTGCGAGGA) amplified a sequence of 220 bp. and VIRI900FP7 (TACGCTCCAGGCTACCACTT) VIRI900RP7 (GAGATGAGCTCCTTGCTGCT) amplified a sequence of 900 bp. which was specific to T. harzianum and T. viride, respectively. The specificity of the marker when tested against six isolates of Trichoderma species showed a specific band of 220 bp. only in T. harzanium and a specific band of 900 bp. only in T.viride with the optimized PCR parameters. This sequence characterized amplified region (SCAR) marker was sensitive and could detect small quantities of Trichoderma DNA as low as 25 to 30 ng with high efficiency. This marker could also clearly distinguish T. harzianum and T. viride from other isolates of Trichoderma.



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