Scientific Program

Day 1 :

  • Keynote Forum
Location: Orlando, USA

Session Introduction

Laying Du

Lindsley F. Kimball Research Institute New York Blood Centre, USA.

Title: MERS-CoV receptor-binding domain-based vaccines and therapeutic antibodies
Speaker
Biography:

Dr.Laying Du has over 10 years’ experience in designing and developing effective vaccines against emerging infectious viruses with more than 100 papers in the related research fields. The research focuses of Dr. Du are on 1) the development of vaccines and therapeutic antibodies for the prevention and treatment of emerging infectious diseases caused by influenza viruses, coronaviruses, including Middle East respiratory syndrome coronavirus (MERS-CoV) and severe acute respiratory syndrome coronavirus (SARS-CoV), as well as blood transfusion-associated virus, including Zika virus (ZIKV); and 2) the study of pathogenic mechanisms of these viruses, based on which to design novel vaccines and therapeutic antibodies.

Abstract:

Middle East respiratory syndrome (MERS)-coronavirus (MERS-CoV), an emerging infectious coronavirus, was first identified in 2012, and continues to threat public health with a high mortality rate (~36%), reinforcing the urgency to develop safe, effective, and innovative vaccines and therapeutic antibodies to prevent and treat MERS-CoV infection. The MERS-CoV spike (S) protein mediates virus binding to its receptor dipeptidyl peptidase 4 (DPP4) via the receptor-binding domain (RBD) and subsequent virus-cell membrane fusion, thus serving as a key vaccine and therapeutic target. In this study, we have identified the RBD of MERS-CoV S protein as a critical neutralizing domain fragment in the development of MERS vaccines and therapeutic antibodies. Our data demonstrate that recombinant RBD vaccines induce potent and broad-spectrum neutralizing antibodies against MERS-CoV of different circulating human and camel strains, as well as antibody escape MERS-CoV mutants. In particular, a variant form of RBD vaccines with an unfavorable epitope masked in the RBD significantly enhances efficacy, completely protecting human transgenic mice from lethal MERS-CoV challenge. In addition, we have found that MERS-CoV RBD-targeting neutralizing antibodies recognize highly conserved RBD epitopes and significantly block RBD binding to viral receptor DPP4, potently cross-neutralizing MERS-CoV at divergent strains. Importantly, single-dose, prophylactic and therapeutic treatments with these neutralizing antibodies protect human transgenic mice from MERS-CoV infection with 100% survival rate. Collectively, our study demonstrates the importance and feasibility for further development of RBD-based subunit vaccines and therapeutic antibodies against MERS-CoV.

  • Virology | Infectious Diseases
Speaker
Biography:

Dr Trang has his expertise in clinical medical microbiology. He has been working in a hospital in Hanoi for 13 years. Since 2003, after the SARS outbreak, avian influenza H5N1 occurred in North Vietnam, he proposed a detailed plan to receive supports from experts from the Oxford University Clinical Research Unit (OUCRU. He gained experience and expanded co-operation with other reputable organizations around the world in research and training. Consequently, he has been studied in BK polyomavirus in the University of Sydney since 2015. He was author and co-author of some recent following papers. 1.Trang VD, Rockett R, Jeoffreys N, Trung NV, Hai An HP, Kok J, Dwyer DE. 2017. BK polyomavirus: a review of the virology, pathogenesis, clinical and laboratory features, and treatment. Future Virol. 2.Van TD, Dinh Q-D, Vu PD, Nguyen TV, Van Pham C, Dao TT, Phung CD, Hoang HTT, Tang NT, Do NT. 2014. Antibiotic susceptibility and molecular epidemiology of Acinetobacter calcoaceticus–baumannii complex strains isolated from a referral hospital in northern Vietnam. Journal of global antimicrobial resistance 2:318-321. 3. Le MT, Wertheim HF, Nguyen HD, Taylor W, Hoang PV, Vuong CD, Nguyen HL, Nguyen HH, Nguyen TQ, Nguyen TV. 2008. Influenza A H5N1 clade 2.3. 4 virus with a different antiviral susceptibility profile replaced clade 1 virus in humans in northern Vietnam. PLoS One 3:e3339.

Abstract:

BK polyomavirus (BK virus or BKPyV) is a circular double-stranded DNA virus with an approximately 5100 bp genome. BKPyV is opportunistic pathogen related to several human diseases under immunosuppressed status. It affects mostly renal and bone marrow transplant patients, and may cause BKPyV-associated nephropathy (BKVAN) occurring in 1-10% of renal transplant recipients. It is divided into four major genotypes (BKPyV I-IV). However, there are limitations of a 287-bp VP1 region to sub-classify genotype subgroups. The aim of this study was to detect the distribution of BKPyV genotypes and subgroups in Australian and Vietnamese transplant recipients (TRs), and to compare the improvement of phylogenetic analysis based on whole genome sequencing (WGS) rather than 287-bp VP1 region sequencing. BKPyV DNA-positive blood and urine samples were analyzed from Australian and Vietnamese TRs. Primer-directed rolling circle amplification method was used to enable sufficient polyomavirus-specific WGS reads for accurate assessment of the BKPyV whole genome. Advances in next-generation sequencing (NGS) technologies is an effective approach to explore genetic diversity over the BKPyV complete genome. NGS was performed on a BKPyV DNA-enriched WGS directly from clinical samples. Phylogenetic analysis of BKPyV based on WGS and VP1 region, four major genotypes I, II, III and IV were detected in Australia, whereas only genotypes I and IV were found in Vietnam. Genotype I was classified into four subgroups I-a, I-b1, I-b2 and I-c in Australia, while subgroup I-b1 was detected in Vietnam. Within genotype IV, it was classified into IV-a2 and IV-c2 subgroups in Australian TRs, and three subgroups IV-a1, IV-a2 and IV-c1 in Vietnamese RTRs. However, the 287-bp VP1 region poorly differentiated subgroups in genotype IV. Particularly subgroups IV-c1 and IV-c2 compared to WGS phylogeny. Phylogenetic analysis based on the WGS is more accurate than the 287-bp VP1 region in classifying BKPyV genotypes/subgroups.

Speaker
Biography:

Dr. Ola El Madani has completed her Bachelor of Medicine and Bachelor of Surgery from Medical school, Al Azhar University, Cairo ,Egypt. She acquired her Master’s degree in Microbiology in 1986 and Phd doctorate degree in Virology in the year 1989. She currently works as an Assistant Professor and Director of Virology lab in Student health hospital, Mansoura University, Mansoura, Egypt.

Abstract:

HCV affects approximately 185 million people around the world. HCV infections are one of the few infections causing psychiatric disorders. HCV load in the human body per se induces an inflammatory process in the central nervous system. Patients suffering from Chronic HCV also endorsed comorbid psychiatric disorders like depression, sleep disorders, anxiety and bipolar disorder. The conventional gold standard treatment of HCV up till 2011 was the combination of Pegylated interferon and Ribavirin. This treatment was reported to interfere with the hypothalamic pituitary adrenal axis. Causing depression in about 45-60% of cases, anxiety, sleep disturbance, anger, irritability in 75% of cases, mania, sexual impairment and suicidal ideations. In 2011 HCV treatment has evolved from interferon-based treatment to direct-acting antiviral (DAA) Extensive literature review showed that psychiatric symptoms did not increase during treatment with DAA medication. Psychiatric symptoms even decreased with the DAA treatment. In this study we conducted a Medline and PUBMED search. Keywords that were used in the search included. Interferon alpha, Ribavirin, HCV, Sofosbuvir.

Conclusion: For the treatment of Chronic HCV.DAA was found to be more tolerable than conventional antiviral medication. DAA does not induce psychiatric disorders and does not exacerbate underlying psychiatric disorders.

  • Global Spread of Virus

Session Introduction

Ely John Karimela

Nusa Utara State Polytechnic, Indonesia.

Title: The existence of Microbes in Pinekuhe Smoke Fish Marketed in Sangihe Islands District
Speaker
Biography:

Abstract:

It is one of the resultant product which prepared by the smoke fisheries have owned by the fisherman in Regency of the Sangihe Island known as Pinekuhe. In researching of this aim is to know the higher contamination of microbe on the smoke fish pinekuhe it was the product and prepared by the fisherman in the Regency of the Sangihe island. In observing of the declining mutual product of smoke fish pinekuhe, it had been teachable through the TPC, have a total of mold or shipworm from the total of Staphylococcus sp. the result of the experiment had been indicated value of TPC, to the Processor A, and have a value of 1.3 x 104 CFU/g at the same level in TPC. The Processor B have value of 2.6 x 104 CFU/g in TPC. In Towoe market the product of TPC have own a value of 6.9 x 104 in TPC. Even in the Petta market have value of 1.2 x 104 CFU/g in TPC. From the total result of observing Staphylococcus sp. to the Processor A and Processor B have a product value of zero. Towoe market has a total product of Staphylococcus. and have own total values of 1.1 x 104 TVC/g, at the same level. While in Petta Market have a total value of 1.2 x 104 TVC/g in Staphylococcus sp. In analyzing total product of Fungi to the Processor A and Processor B have product value of zero. Towoe market has own total value of 1.3x 105 CFU/g in Fungi, on the same level. Even in Petta market a total value of 5.9 x 104 CFU/g in fungi in the same level. Therefore, they have a total bacterium as from the Processor A and B. Towoe Market and Petta have an overall total of fungi from the Processor A and B and have total staphylococcus sp. in Processor A and B are still following conditional system while the total of fungi and the total of Staphylococcus sp. To the Towoe Market and Petta Market have much more total conditions from the standard National Indonesia which have concerning limits of dirtiness microbe to the smoke fish.

  • Development of Antiviral | Host Defences | Virus Vaccines

Session Introduction

Hina Ashraf

University College of Veterinary and Animal Sciences, Pakistan.

Title: Antiviral activities from selected Cholistani plants against common viral diseases of poultry birds
Speaker
Biography:

Miss Hina Ashraf has expertise in Plant Biotechnology and Medicinal plants. She is associated as a faculty member in the department of Botany with Govt. Sadiq College Women University Bahawalpur Pakistan since 2014. She has supervised many research students and also has played an active role in winning  projects for her department. She has expertise in the evaluation of anti bacterial, anti viral , anti oxidant anti diabetic and many other biological activities of Cholistani medicinal plants.      

Abstract:

The idea of herbal medicine is getting popular day by day and demand for herbal products is continuously rising due to its usefulness, limited side effects and cost-effectiveness. Cholistani plants are reported as a rich source of antibacterial, antifungal, antiprotozoal, antioxidant and anticancer agents but their antiviral potentials were not explored before. The current study was designed to asses the antiviral capabilities of selected Cholistani plants. The whole plants were collected and authenticated by a taxonomist. Plants were shade dried and powdered form was used to make extracts with methanol, n-hexane, ethyl acetate and n-butanol. These extracts were concentrated by using a rotary evaporator and 0.5g each precipitate was dissolved in 1mL of the respective solvent to make final extract. All of these fractions were tested for their antiviral potentials through 9 to 11 days old chick embryonated eggs. HA test was performed to asses’ antiviral potential of each fraction against Avian Influenza virus H9N2 strain, New Castle Disease virus Lasota strain and Infectious bronchitis virus similarly, IHA test was used quantification of Infectious bursal disease virus. The order of antiviral activities of Cholistani plants was NDV>IBV>IBDV>AIV. In terms of antiviral activity from different fractions, the order of activity was n-butanol>ethyl acetate>methanol>n-hexane. The extracts of medicinal plants A. Aspera, N. procumbus antidotale, . compressa and S. fruticose were effective against all four poultry viruses. The low IC50values of these plant extracts confirm the very high antiviral potentials of these plants against above-mentioned poultry viruses. It is worth to mention that all extracts of A. aspera were positive against IBDV, which has no treatment in the market so far.

Biography:

Dr. Yacob Mithai is a practicing physician in the field of healthcare in the state of Kerala in India for the last 29 years. His interest is spread across the fever, inflammation and back pain. he has printed and published nine books in the related subjects. he has written hundreds of articles in various magazines and presented 9 research papers in Indian Science Congress in the years 2008, 2009 & 2010,2016, 2017. And also 2 papers have been selected by Indian science congress for the coming 2018.

Abstract:

According to the facts of physics, if temperature increases, thermal expansion of an object is positive it will expand and with decrease of temperature it will shrink. Pressure will increase due to increase of temperature.On the contrary, during fever we can see blood vessels and skin are shrunk, pressure decreases, body shivers, sleep increases, motion decreases, inflammation increases, body pain increases, blood circulation decreases, dislike cold substances etc,. In fever, the firing rate of Warm sensitive neurons decreases, and the firing rate of Cold sensitive neurons increase. At the same time if we apply hotness from outside by thermal bag or if we drink hot water, our body acts according to the Facts of Physics- increase of temperature pressure will also increase, expands blood vessels and skin, body sweats, motion will increase, inflammation will decrease , body pain will decrease, blood circulation will increase, like cold substances etc.. During fever, why our body acts against Facts of Physics? We will get a clear answer if we find out the purpose of the fever. No medical books clarify this. During fever, if the temperature of fever is not a surplus temperature or if it is not supposed to be eliminated from the body, the shrinking of skin and blood vessels, shivering of body, dislike towards cold substances etc. are a protective covering of the body to increase blood circulation to important organs of the body it is not against the facts of physics.

Day 2 :

  • Keynote Forum
Speaker
Biography:

Dr Yusuf Hussaini have spent more than a decade teaching and supervising projects in the fields such as Virology, Immunology, Molecular Biology and Microbial Genetics. During this time I have worked on a number of researches to formulate and improvise methods of navigating through the deplorable economic problems in my Nigeria to achieve maximum health benefits using minimal resources. In all these years I have worked closely with experts to connect hypotheses to experiment. Over the coming decade, I will continue to work on important viral problems comparing efficiencies and effectiveness of different methods and approaches to viral diseases cures.

Abstract:

Varicella-zoster virus (VZV) is the causative agent of Chicken pox (varicella) and shingles (zoster).Varicella is a sudden onset, very contagious disease that is characterized by a widespread, blister-like rash.It typically infects children in temperate regions; children, mostly adolescents’ and adults are infected in tropical region. This cross-sectional study evaluated the seroprevalence of Varicella-zoster virus (VZV) immunoglobulins gamma (IgG) and mu (IgM) by Enzyme Linked Immunosorbent Assay (ELISA). Serum samples obtained from respondents were tested using a whole-virus IgG and IgM VZV-specific commercial enzyme-linked Immunosorbent assay (ELISA) kits. Respondents’ demographic data were obtained using questionnaires. Ethical consent was sought from the appropriate authority for both minors and adults. A total of seven hundred and fifty (750) respondents domiciled in North-central Nigeria had their serum samples analyzed. The overall seroprevalence of 71.7 % and 26.7 % were obtained for IgG and IgM antibodies respectively. The highest Seroprevalence of 86.0% for IgG was recorded in Niger state while the highest seroprevalence of 33.6% for IgM was recorded in the federal Capital territory Abuja. Seroprevalence of VZV IgG by age recorded the highest rate of 78.1% among those 41 years and above while for those 10 years and below the lowest rate was 65.4 %. The prevalence rate for male and females were 71.0 % and 72.5 % respectively. These results demonstrate a high prevalence of varicella among the population studied. We recommend instituting improved education regarding varicella among communities and increasing vaccination program or routine testing for serum varicella antibody to prevent VZV-related morbidity and mortality, particularly in adolescents and adult.

  • Virology | Diagnosis Of Viral infections | Viral Infections
Speaker
Biography:

Assistant Prof Elvis Asare-Bediako has published various research papers and articles in reputed journals and has other achievements in the related studies. He extended his valuable service towards the scientific community with his extensive research work.  Pest management; Pest control; Crop protection practices; Agricultural knowledge and information systems; Farmers’ perceptions; Hazard; Pesticide policy

Experience: Department of Crop Science, School of Agriculture, University of Cape Coast, Ghana | Biotechnology and Nuclear Agriculture Research Institute Ghana | Atomic Energy Commission, Accra, Ghana | Department of Agricultural Economics and Extension, School of Agriculture, University of Cape Coast, Ghana | CSIR-Crop Research Institute, Kumasi, Ghana

 

Abstract:

Maize streak disease (MSD) is the most serious biotic constraint to maize production in sub-Saharan Africa, threatening food security and incomes of farmers in the region. Field trials were established at Ohawu, Kpeve and Nkwanta representing coastal savannah, forest and transition ecozones respectively of the Volta region, during 2015 minor season to evaluate 16 maize genotypes for high yielding and resistance to maize streak virus (MSV) infection. The fields were laid out in a randomised complete block design (RCBD) with four replications. The study revealed significant varietal, locational and genotype x location interaction effect (P<0.05) on mean MSD severity scores and grain yields. Mean MSD severity scores were highest at Nkwanta, followed by Kpeve and the lowest at Ohawu. The improved varieties namely Abontem, Aburohemaa, Akposoe, Dorke, Etubi, Honampa, Omankwa, Obatanpa, PAN 12, and the breeding lines CRI001, CRI002, CRI003, CRI004, CRI005 and CRI006 which exhibited mild MSD symptoms had higher yields than Dzinu-Eve (a local cultivar). However, the study revealed significant genotype x location interaction effect, with Akposoe, Etubi, Honampa and PAN 12 showing higher MSD symptom severity scores at Kpeve than at Nkwanta and Ohawu, whereas the other 12 maize genotypes had higher mean MSD severity scores at Nkwanta than Kpeve and Ohawu.  It is suggested that genotypes that showed mild symptoms of MSD and also gave high grain yields should be further evaluated and released as a varieties for the locations where they performed better.

 

Speaker
Biography:

Dr Mirza Imran Shahzad is involved in active research since 2001. He did PhD in joint venture of University of California Davis and PMAS Arid Agriculture University Rawalpindi, Pakistan. He has produced more than 45 research articles in world renowned international journals. He has produced more than 25 post graduate students. His area of interest is DNA vaccines against Tuberculosis and Bird Flu virus. Similarly, his research group is also working on medicinal plants of Cholistan region of Pakistan. His lab is well equipped to perform antibacterial, antiviral, antioxidant and many more biological activities. Dr Shahzad’s lab has many international collaborations with different countries including USA and UK. He is travelled more than 10 countries in the world. 

Abstract:

Cholistan desert is a rich source of medicinal plants. These plants have been reported for many biological and pharmacological activities including antibacterial, antifungal, anti diabetic, antiviral etc. but very few chemistry based approaches are available to classify principal active agent(s). The current study was designed to evaluate the antibacterial and antiviral potentials of different extracts of Cholistani plants against five common disease causing bacteria and bird flu virus isolated from Bahawalpur region. Aqueous, n-hexane, methanol, n-butanol and ethanol extracts were made and tested against each bacterium through disc diffusion method and in ovo antiviral assay against bird flu virus. IC50 and MIC of each positive extract was calculated. In antibacterial study, the results indicate that aqueous extracts have produced greater zones of inhibition as compared to other extracts. The overall trend of activity was aqueous>n-hexane>methanol>n-butanol>ethanol. In terms of bacteria, these extracts were effective in given order Klebsiella pneumonia> Proteus vulgaris> Staphylococcus aureus> Escherichia coli> Pseudomonas aeuriginosa. In antiviral studies, the order of activity of extracts were n-butanol>methanol>ethanol>aqueous>n-hexane. Selected plant extracts were subjected to TLC and HPLC analysis. The study concludes that Cholistani plants are rich source of antibacterial and antiviral agents and according to their nature(s), these compounds are soluble in different solvents.

Speaker
Biography:

Dr. Fares Wasfi has done his PhD and currently working more on the Virology Department. He is a scholar with  89 Research items and Massive project on Midgut bacterial flora and innate immunity in sand flies.

Abstract:

Toscana virus (TOSV), a sandfly-borne virus, is transmitted to human by the bite of sandfly of the subgenus Larroussius, mainly Phlebotomus perniciosus and Phlebotomus perfiliewi species. TOSV infections can lead to the outbreaks of acute meningitis or meningo-encephalitis cases. Until now, the epidemiological status of TOSV infections remains largely unknown. Here, we performed a Bayesian Markov Chain Monte Carlo analysis with strict and relaxed molecular clocks to estimate the demographic history of the TOSV infection in a series of isolates sampled between 1971 and 2015. The present results showed that the tmrca of the TOSV in the Mediterranean basin occurs around 1489 (927-1795) with a tmrca in Tunisia around 1981 (1928-2004). Bayesian skyline plot revealed a short decrease in the effective number of infections around 1975 with a net increase during the last 40 years in North and South Mediterranean countries. The phylogeography of TOSV showed that all circulating lineages in the Mediterranean basin and in Tunisia spread from Italy. The present study contributed to reveal the time of origin and patterns of geographical dispersal of the different isolates in the Mediterranean basin, particularly in Tunisia.

  • Plant and Animal Virology | Virus Vaccines | Emerging Viruses
Speaker
Biography:

Ali ilter Akdag pursuing his Ph.D. from Jamia Hamdard University for three years. His research area is molecular virology and vaccinology. He works on acute gastroenteritis viruses, like rotavirus, adenovirus, astrovirus, norovirus, and sapovirus. He has completed MSc(Zoology) from Osmania University 2013-2015 in Hyderabad, India. He has many certificates and awards. He has actively participated conferences national and international.

Abstract:

Background: This ( rotavirus, astrovirus, and adenovirus) viruses are responsible for diarrhea in children, basically those have near about <5 years old. Detection of the entire virus is crucially important to the development of the effective cure. This study aimed to determine the prevalence of common these viruses in children <5 years old presented with diarrhea to the from Lala Lajpat Rai Memorial Medical College (LLRM) centre(Meerut) North India, India

Methods: Total 312 fecal samples were collected from diarrheal children duration 3 years: in year 2014 (n = 118), 2015 (n = 128) and 2016 (n = 66) ,<5 years of age who presented with acute diarrhea at the Lala Lajpat Rai Memorial Medical College (LLRM) centre(Meerut) North India, India. All samples were the first detection by EIA/RT-PCR for rotaviruses, adenovirus and astrovirus. Results: In 312 samples from children with acute diarrhea in sample viral agent was found, rotavirus A was the most frequent virus identified (57 cases; 18.2%), followed by Astrovirus in 28 cases (8.9%), adenovirus in 21 cases (6.7%). Mixed infections were found in 14 cases, all of which presented with acute diarrhea (14/312; 4.48%). Conclusions: These viruses are a major cause of diarrhea in children <5 years old in North India. Rotavirus A is the most common etiological agent, follow by astrovirus. This surveillance is important to vaccine development of the entire population.

Speaker
Biography:

Dr. Rasha Elkady is a Co-chair of the MPPA ECP Committee and works as an Assistant Professor at University of Missouri at Columbia. She is also a member of 5 Professional Psychiatry Societies. She has around 9 publications and is currently working on Absence Seizures causing Visual hallucination in Children.

Abstract:

HCV affects approximately 185 million people around the world. HCV infections is one of the few infections causing psychiatric disorders. HCV load in the human body per se induces an inflammatory process in the central nervous system. Patients suffering from Chronic HCV also endorsed comorbid psychiatric disorders like depression, sleep disorders, anxiety and bipolar disorder.

The conventional gold standard treatment of HCV up till 2011 was the combination of Pegylated interferon and Ribavirin. This treatment was reported to interfere with the hypothalamic pituitary adrenal axis. Causing depression in about 45-60% of cases, anxiety, sleep disturbance, anger, irritability in 75% of cases, mania, sexual impairment and suicidal ideations.

In 2011 HCV treatment has evolved from interferon-based treatment to direct acting antiviral (DAA) Extensive literature review showed that psychiatric symptoms did not increase during treatment with DAA medication. Psychiatric symptoms even decreased with the DAA treatment.

In this study we conducted a Medline and PUBMED search. Key words that were used in search included. Interferon alpha, Ribavirin, HCV, Sofosbuvir.

 

 

Speaker
Biography:

Chakradhar Pokkula is a Assistant professor in virology at Success Degree College of Commerce & Science, Telangana university. He received master’s degree in Infection Biology from Uppsala university, Sweden and Bachelor’s degree in Microbiology from Osmania university, India. He worked as a research assistant in Uppsala academic hospital where he worked on developing Antivirals and resistance mechanism. His research interests include development of antiviral agents for dengue virus, Zika and vaccines for emerging viruses.

Abstract:

There are arrays of in vitro assays to quantify the activity of HIV-1 reverse transcriptase (HIV-1 RT). These assays utilize either chemically customized/labelled nucleotides, or TaqMan probes, or radiolabeled nucleotides/primers. Although several real-time PCR assays exist commercially for measuring the RT activity, which are usually used for quantifying the viral titres, these assays are not optimized for measuring the inhibitory concentrations (IC50) of HIV-1 RT inhibitors. Moreover, a recently established inorganic pyrophosphate-coupled enzyme assay cannot be employed for studying nonphosphorylated nucleoside reverse transcriptase inhibitors (NRTIs). In the present study, we have developed a novel one-step assay with native nucleotide substrates and SYBR Green II dye to determine IC50 values of triphosphorylated NRTIs against HIV-1 RT. Using exact batches of wild-type and mutant RT, and triphosphorylated NRTIs, we showed that our method gave IC50 values for inhibitors similar to that of an earlier published colorimetric assay with BrdUTP substrate (CABS). Our assay should be suitable for high-throughput screening of antiretroviral drugs and could also be suitable for studying drug resistance profiles. Additionally, we also used our assay to study inhibition by AZT in its nonphosphorylated form by supplementing the reaction mixture with necessary kinases and ATP.