Bassine Science Building, 103
DegreesRockefeller University, Ph.D.
Brandeis University, B.A.
ExpertiseLinear-After-The-Exponential (LATE)-PCR and Its Allied Technologies for highly informative, highly accurate DNA amplification and application of these technologies for design and construction of diagnostic assays in many fields.
ProfileScientists in our laboratory have overcome the inherent limitations of symmetric PCR by developing an advanced form of asymmetric PCR, known as LATE-PCR. Symmetric PCR has been the dominant method of DNA amplification for the last 20 years. Each amplicon in such assays is generated by use of two primers of equal concentration and equal Tm (melting temperature) and is therefore double-stranded. Amplification proceeds exponentially, but then slows down and reaches plateau in a stochastic manner. Thus, although such reactions are well known and are often robust, they nevertheless suffer from limitations which are particularly evident in samples containing low numbers of initial targets. These limitations include:
* Design difficulties, particularly for multiplexing
* Reduced efficiency and sensitivity due to mis-priming
* Variability among replicate reactions, particularly at end-point
* Limited availability of fluorescent colors
* Limitations in probe design
* Requirement for hot-start enzymes
LATE-PCR and Its Allied Technologies, including PrimeSafe, Lights-On/Light-Off Probes, and Dilute-N-Go Sequencing overcomes all of the above problems and makes it possible to construct highly-informative single tube assays using fluroescent probe in several colors. These assays make it possible to simultaneously scan multiple targets for sequence variations (i.e Virtual Sequencing in a closed-tube). Over the past 15 years we have worked with several large international diagnostic companies and have built numerous assays in the fields of human and animal infectious diseases, human cancers, forensics, drug side effects, species identification, prenatal and pre-implantation genetic diagnosis. We have had over 10 million dollars of funding and have built a large patent portfolio.
|BIOL||154b||Project Lab: Barcoding and Virtual Barcoding of Species|
|BIOL||350a||Graduate Student Research Seminar|
|BIOL||350b||Graduate Student Research Seminar|
|BIOL||350d||Graduate Student Research Seminar|
|BISC||2b||Genes and the Human Story|
Awards and Honors
2009-2011 Sigma Xi Society - Distinguished Lecturer (2009 - 2011)
Feldman Brothers Award for Course Development in human reproduction and population explosion (1999)
Helen Hay Whitney Postdoctoral Fellowship (1973 - 1975)
A. Osborne, J.A. Sanchez, L. Wangh, K.C. Hayes. The Mitigation of Cytotoxic and Genotoxic Effects of Drugs on Mitochondrial DNA. Proc. of Targeting Mitochondrial Dysfunction & Toxicity. Boston, MA: Cambridge Healthtech Institute, 2015.
Alexandra Over, Wangh, Lawrence J. ThermaStop™ Hot-Start Reagent: Properties and Benefits. Proc. of qPCR and NGS 2015 Symposium. Freising, Germany: 2015.
Kenneth E. Pierce and Lawrence J. Wangh. "Low-concentration Initiator Primers improve the amplification of gene targets with high sequence variability.." Methods in Molecular Biology. Ed. Basu, Chhandak. Springer Science+Business Media New York 2015, 2015. Chapter 5.
Kenneth Pierce and Lwrence J Wangh. Opening New Dimensions in Diagnostics with Hybridization Probes. Proc. of qPCR and NGS 2015 Symposium. San Francisco, CA: Select Bioscience, 2015.
Osborne AE, Sanchez JA, Wangh LJ, Sambanthamurthi R, Hayes KC .. "Oxidative Damage is not a Major Contributor to AZT-Induced Mitochondrial Mutations." J AIDS Clin Res 6:. 441 (2015): doi:10.4172/2155- 6113.1000441.
A. Osborne, J.A. Sanchez, L Wangh (Poster Presentation). Palm fruit juice mitigates the cytotoxic and genotoxic effects of AZT and isoniazid, key drugs in the treatment of HIV and tuberculosis. Proc. of International Union Against Tuberculosis and Lung Disease, 45 World Congress. Barcelona, Spain: 2014.
J Rice, L Wangh, R M Warren, M Whitfield, S Marras, B Kreiswirth (Oral Presentation). Single tube two-colour analysis of all allelic variants of the pncA gene using LATE-PCR and allied technologies. Proc. of International Union Against Tuberculosis and Lung Disease, 45th World Congress. Barcelona, Spain: 2014.
J. Rice, K. Pierce, L. Wangh. "Single Tube Single-Color Analysis of Rifampicin and Isoniazid Allelic Variants in MDR-TB." International Congress on Infectious Disearse. Cape Town, South Africa. April, 2014.
John Rice, Kenneth Pierce, Lawrence Wangh (Oral Presentation). A Single-Tube Single-Color Assay for All Allelic Variants of the first line gene targets inhA, katG, and rpoB that cause MDR-TB. Proc. of International Union Against Tuberculosis and Lung Disease, 45th World Congress. Barcelona, Spain: 2014.
Kenneth E. Pierce and Lawrence J. Wangh. "Opening New Dimensions in Diagnostics with Hybridization Probes." Select Bioscience, Genomics 2014. San Francisco. April 2014.
Kenneth E. Pierce and Lawrence J. Wangh. "Rapid detection and identification of hepatitis C virus (HCV) sequences using mismatch-tolerant hybridization probes: A general method for analysis of sequence variation." BioTechniques Septembe 2013. doi 102144/000114076 (2014): 125-132.
L Wangh, A Osborne, K.C. Hayes. Mitochondrial mutational load in diabetes: toward a better understanding of diabetes in tuberculosis. Proc. of International Union Against Tuberculosis and Lung Disease, 45 World Congress. Barcelona, Spain: 2014.
M De Vos, J Rice, L Rice, B Kreiswirth, E Streicher, P Van Helden, R M Warren, L Wangh (Poster Presentation). Detecting resistance to nine anti-tuberculosis drugs in a single reaction tube. Proc. of International Union Against Tuberculosis and Lung Disease, 45 World Conference. Barcelona, Spain: 2014.
M De Vos, J Rice, P Van Helden, R M Warren, L. Wangh (Poster Presentation). Construction of a three colour single-tube assay for resistance to first, second and third line antibiotics. Proc. of International Union Angainst Tuberculosis and Lung Diseases, 45 World Conference. Barcelona, Spain: 2014.
Osborne AE, Sanchez JA, Solomon M, Stopa A, Wangh LJ, et al.. "Palm Fruit Juice Mitigates AZT Mitochondrial Genotoxicity and Dose-Dependent Cytotoxicity.." J AIDS Clin Res 5:. 400 (2014).
Rice, J., Rice, L., Wangh, L., Kurpina, N. Kreiswirth, B., De Vos, M., Warren, R.. "Single-Tube Multiplex Assay for M(X)DR-TB." North American Region of the International Union Against Tuberculosis and Lung Disease. 18th Annual Conference, Boston. February, 2014.
Shana M. Tetrault, John E. Rice, Lawrence J. Wangh and J. Aquiles Sanchez. "Single-Tube Mutation Scanning of the Epidermal Growth Factor Receptor Gene using Multiplex LATE-PCR and Lights-On/Lights-Off Probes." Journal of Molecular Biomarkers & Diagnosis 5. 175 (2014): doi:10.4172/2155-9929.1000175.
Wangh, Lawrence J. "A Single-Tube Single-Color Assay for All Allelic Variants of inhA, katG, and rpoB, plus two Internal Controls." Hain Lifesciences. London, England. January, 2014.
Wangh, Lawrence J. "Evaluation & Validation of a Highly Multiplexed LATE-PCR Single-Tube Assay for M(X)DR-TB." International Congress of Infectious Disease. Cape Town, South Africa. April, 2014.
Wangh, Lawrence J. "Virtual Barcoding." Cambridge Healthtech Institute, Tri Conference. San Francisco, CA. February, 2014.
Yanwei Jia, J. Aquiles Sanchez, and Lawrence J. Wangh. "Kinetic Hairpin Oligonucleotide Blockers for Selective Amplification of Rare Mutations." SCIENTIFIC REPORTS 4. 5921 (2014): DOI: 10.1038/srep05921.
Adam E. Osborne*, John E. Rice, J Aquiles Sanchez and Lawrence J. Wangh. "AZT Treatment Increases mtDNA Mutations in HepG2 and CCD-1112Sk Cells." J AIDS Clin Res Volume 4. 11 (2013): doi: 10.4172/2155-6113.1000250.
Adam Osborne and Lawrence Wangh. "Analysis of Heteroplasmy Among Mitochondrial Genomes at the Single Molecule Level using LATE-PCR and Allied Technologies." Select Biosciences. Boston, MA. May 2013.
Arthur Reis and Lawrence Wangh. "Sepsis, MRSA, VAP: Molecular Pathogen and Antibiotic Resistance Detection in a Single Tube Multiplex LATE-PCR Assay." Select Biosciences. Boston, MA. May 2013.
J. Aquiles Sanchez and Lawrence J. Wangh. "Single-tube Mutation Scanning of Epidermal Growth Factor Receptor Gene using Multiplex LATE-PCR and Lights-On/Lights-Off Probes." Select Biosciences. Boston, MA. May 2013.
J. Rice, L. Rice, L. Wangh, N. Kurpina, B. Kreiswirt, N. Casali, F. Drobniewski, M. De Vos, R. Warren. "A Four Color, Highly Multiplexed, Single-Tube, Quantitative End-Point Assay for M(X)DR-TB using LATE-PCR and Allied Technologies." European Society of Mycobacteria. Florence Italy. July 2013.
Kenneth E. Pierce, Harald Peter, Till T. Bachman, Carmelo Volpe, Rohit Mistry, John E. Rice, and Lawrence J. Wangh. "Rapid Detection of TEM-type Extended Spectrum Beta Lactamase (ESBL) mutations using multiple Lights-On-Lights-Off Probes with single-stranded DNA amplification." Journal of Molecular Diagnostics Vol. 15, No. 3, May 2013. No. 3 (2013): http://dx.doi.org/10.1016/j.jmoldx.2013.02.002.
Kenneth Pierce and Lawrence J. Wangh. "Highly Divergent Gene Families are Efficiently Amplified Using Low-concentration Initiator Primers." Select Biosciences. Boston, MA. May 2013.
L.M. Rice, A.H. Reis Jr, R. Mistry, H. Khan, P. Khosla, S. Bharya and L.J. Wangh. "Design and construction of a single tube, quantitative endpoint, LATE-PCR multiplex assay for ventilatorassociated pneumonia." Journal of Applied Microbiology 11 June 2013 (2013): doi:10.1111/jam.12281.
M. de Vos, J. Rice, B. Kreiswirth, E.M. Streicher, P.D. van Helden, R.M. Warren, and L. Wangh. "Evaluation and Validation of a highly-informative LATE-PCR single-tube assay for M(X)DR-TB." European Society of Mycobacteria. Florence, Italy. July 2013.
Osborne AE, Rice JE, Sanchez JA, Wangh LJ. "AZT treatment increases mtDNA mutations in hepG2 and CCD-1112Sk cells.." J AIDS Clin Res 4:. (2013): 250..
Rice, J. and Wangh, Lawrence J. "Perfect PCR: A Quest for Control." Select BioScience. Boston. May 2013.
Wangh, Lawrence J, Webinar, October 2, 2013. How to Achieve Perfect PCR Vial Suppression of Mis-Priming Before, During, and After Amplification. Biotechniques
Wangh, Lawrence J. "Key Note Speaker: Analysis of M(X)DrTB using LATE-PCR with Lights-On/Lights-Off Probes or Lights-Off Only Probes." Select BioSciences. Boston. May 2013.
Wangh, Lawrence J. "LATE-PCR and Allied Technologies Maximum Information from a Single Tube M(X)DR TB Assay." Foundation for Innovative Diagnostics. Geneva, Switzerland. November, 2013.
Yanwei Jia, Pui-In Mak, Conner Massey, Rui P. Martins, and Lawrence J Wangh. "Construction of a microfluidic chip, using dried-down reagents, for LATE-PCR amplification and detection of single-stranded DNA." Lab on a Chip 13. DO1:10.1039/c3151049b (2013): 4635.
Harald Peter, Grace Henihan, Kenneth E. Pierce, Kate Templeton, Rohit Mistry, Carmelo Volpe, Lawrence J. Wangh, Till T. Bachmann. "Taking the Bull by the Horns – A new High Content Molecular Assay Addressing Complex ESBL Detection." European Society of Clinical Microbiology and Infectious Disease, March 2012. London, England
John E. Rice, Arthur H. Reis, Jr., Lisa M. Rice, Rachel K. Carver-Brown, Shana Augustin Tetrault & Lawrence J. Wangh. "Fluorescent Signatures for Variable DNA Sequences." Nucleic Acid Research NAR 1-10, doi:10.1093/nar/gks731 (Featured Article). (2012).
Kenneth E. Pierce and Lawrence J. Wangh. "Rapid detection and sequence analysis of Hepatitis C Virus using a novel molecular assay with mismatch-tolerant hybridization probes." CFAR. Harvard Medical School. April 2012.
Kenneth E. Pierce, Huma Khan, Rohit Mistry, Simon D. Goldenberg, Gary L. French, and Lawrence J. Wangh. "RAPID DETECTION OF SEQUENCE VARIATION IN CLOSTRIUM DIFFICILE GENES USING LATE-PCR WITH MULTIPLE MISMATCH-TOLERANT HYBRIDIZATION PROBES." Journal of Clinical Microbiology Journal of Microbiological Methods, 91 (2012) 269–275.. (2012).
L. Rice, J. Rice, S. Fishbein, L. Wangh, N. Kurpina, B. Kreiswirth, N. Casali, F. Drobniewski, EM. Streicher, and R. Warren. "Construction and Evaluation of an M(X)DR-TB Multiplex Assay using LATE-PCR & Thermalight™ Probes." 2012 New England TB Symposium. Broad Institute, Cambridge MA. June 29, 2012.
L. Wangh, J. Rice, L. Rahman, B. Ronish, S. Fishbein, N. Kurpina, B. Kreiswirth, N. Casali, F. Drobniewski, EM. Streicher, and R. Warren. "Evaluation of an M(X)DR-TB Multiplex Assay using LATE-PCR & Thermalight™ Probes." Keystone Symposium: Drug Resistance and Persistance in Tuberculosis. Kampala Uganda. May 13-18, 2012.
L. Wangh, J. Rice, L. Rice, S. Fishbein, L. Rahman, B. Ronish, N. Kurpina, B. Kreiswirth N. Casali, F. Drobniewski, and Rohit Mistry. "Two Single-Tube Multiplex Assays for M(X)DR-TB and NTMs using LATE-PCR & Thermalight™ Probes." World TB Day 2012. Boston University Medical School. March 23, 2012.
Lisa M. Rice, Arthur H. Reis, Jr., Bonnie Ronish, Rachel K. Carver-Brown, John Czajka, Nicole Gentile, Gerald Kost, and Lawrence Wangh. "Design of a single-tube, endpoint, linear-after-theexponential- PCR assay for 17 pathogens associated with sepsis." Journal of Applied Microbiology 114,. (2012): 457--469.
N.L. Gentile, A.M. Dillier, G.V. Williams, J. Ackers, A.H. Reis Jr, L.M. Rice, L.J. Wangh, J.W. Czajka and G.J. Kost. "Verification of monoplex and multiplex linear-after-theexponential PCR gene-specific sepsis assays using clinical isolates." Journal of Applied Microbiology 114, 586--594. (2012): 586--594.
Nicola Casali, Agnieszka Broda, John Rice, Rohit Mistry, Carmelo Volpe, Larry Wangh, Jay Lewington and Francis Drobniewski. "A Novel Single-Tube PCR Assay for Rapid Identification of MDR-TB." European Society of Clinical Microbiology and Infectious Diseases, March 2012. London, England
Osborne, Adam E., Wangh, Lawrence J.. "A LATE-PCR Decaplex Assay for the Detection and Sequencing of Mitochondrial DNA Mutations Down to the Single Genome Level." PloS ONE manuscript submitted. (2012).
Rachel K. Carver-Brown, Arthur H. Reis, Jr., Lisa Rice, John W. Czajka and Lawrence J. Wangh. "Design and Construction of a Single-Tube, LATE-PCR, Multiplex Endpoint Assay with Lights-On/Lights-Off Probes for the Detection of Pathogens Associated with Sepsis." Journal of Pathogens Volume 2012,. Article ID 424808, (2012): 10 pages doi:10.1155/2012/424808.
The Wangh Laboratory Site. LATE-PCR.Org. Brandeis University
Wangh, Lawrence J. "Advances in LATE-PCR and Its Allied Technologies." Select BioSciences: Genomics Research - Advances in qPCR. April 19-20, Boston. http://selectbiosciences.com/conferences/agenda.aspx?pid=190&conf=AqPCR2012.
Wangh, Lawrence J. "Advantages of LATE-PCR Technology." (2012): <http://www.bioquicknews.com/node/761>.
Wangh, Lawrence J. "Very Highly Informative Single-Tube Assays for M(X)DR-TB and Non-Tuberculosis Mycobacteria." Select Biosciences: Molecular Diagnostics Europe, Point of Care Diagnostics, May 10-11, 2012. London, England. http://selectbiosciences.com/conferences/agenda.aspx?pid=350&conf=POC2012.
Wangh, Lawrence J. "GEN Tech Tips PCR: Trends and New Directions." (2011): <http://www.genengnews.com/gen-articles/pcr-trends-and-new-directions/3946/>.
Wangh, Lawrence J. "Genetic Engineering News: “Scaling PCR Workflows from Benchtop to Automation.” Webinar, 29 June 2010." (2011)
Wangh, Lawrence J. "Genome Web: http://www.genomeweb.com/dxpgx/smiths-detection-inks-mdx-license-novartis." (2011)
B. Ronish, M. Hakhverdyan, K. Ståhl, S. Belák, N. LeBlanc, L. Wangh. "Design and Verification of a highly reliable Linear-After-The-Exponential PCR assay for the detection of African Swine Fever virus." J. Veterinary Methods http://dx.doi.org/10.1016/j.jviromet.2010.12.003 (2010).
Hartshorn, Cristina, and Wangh, Lawrence J. "One-step RT-LATE-PCR for mRNA and viral RNA detection and quantification. RT-PCR Protocols: Second Edition." Methods in Molecular Biology series. vol. 2 New York, NY: Springer, 2010
Jia, Y., Hartshorn, C., Hartung, O, Wangh, L.J.. "Heat Shock Memory in Preimplantation Mouse Embryos,." . Fertility and Sterility Vol. 93, No. 8, May 15, 2010 (2010).
Jia, Y., Osborne, A., Rice, J.E., and Wangh, L.J.. "Dilute-‘N’-Go Dideoxy Sequencing of All DNA Strands Generated in Multiplex LATE-PCR Assays." Nucleic Acids Research, doi: 10.1093/nar/gkq111 (2010).
Pierce, K., Mistry, R., Reid, S.M., Bharya, S., Dukes, J.P., Hartshorn, C., King., D.P., Wangh, Lawrence J. "Design and optimization of a novel reverse transcription linear-after-the-exponential PCR for the detection of foot-and-mouth disease virus.." Journal of Applied Microbiology doi:10.1111/j.1365-2672.2009.04640.x online. (2010).
Osborne, A., Reis, A., Bach, L., Wangh, L.. "Single-Molecule LATE-PCR Analysis of Human Mitochondrial Genomic Sequence Variations." PLoS One 4(5). (2009).
Hartshorn, C., Anshelevich, A., Jia, Yanwei, and Wangh, L.J.. "Early Onset of Heat-Shock Response in Mouse Embryos Revealed by Quantification of Stress-Inducible hsp70i RNA." Gene Regulation and Systems Biology 1. (2007): 1-9.
Hartshorn, C., Eckert, J.J., Hartung, O., Wangh, L.J.. "Oct4 and Xist RNA gradients in 8-cell embryos revealed by single-cell duplex RT-LATE-PCR." BMC Biotechnology, 7. 87 (2007): doi:10.1186/1472-6750-7-87.
John Rice, J. Aquiles Sanchez, Kenneth E. Pierce, Arthur Reis, and Lawrence J. Wangh. "LATE-PCR Multi-plexing using PrimeSafe® followed by Multi-Dilute-`N'-Go Dideoxy Sequencing." Nature Protocols 2. 10 (2007): 2429-2438..
Pierce, K.E. and Wangh, L.J., ed. Linear-After-The-Exponential Polymerase Chain Reaction and Allied Technologies:. Methods and Protocols, 2007.
Wangh, Lawrence J. Podcast on LATE-PCR. February 22, 2007. Genetic Engineering News,. <http://www.genengnews.com/genCasts.aspx?id=79>.
J. Aquiles Sanchez, Jessica Abramovitz, Jesse J. Salk, Arthur H Reis, John E Rice, Kenneth E Pierce, and Lawrence J. Wangh. "Two-temperature LATE-PCR endpoint genotyping." BMC Biotechnology 6. 44 (2006).
Jesse J. Salk, J Aquiles Sanchez, Kenneth E. Pierce, John E. Rice, Kevin C. Soares, Lawrence J. Wangh. "Direct Amplification of Single-Stranded DNA for Pyrosequencing using Linear-After-The-Exponential (LATE)-PCR." Analytical Biochemistry Volume 353. Issue 1 (2006): 124-132.
Kenneth E. Pierce and Lawrence J. Wangh, Editor: Alan Thornhill /Series Editor: John Walker, ed. LATE-PCR and allied technologies: Real-time detection strategies for rapid, reliable diagnosis from single cells. Humana Press: UK, 2006.
http://www.brandeis.edu/wanghlab/. Website for LATE-PCR. March 2007. Brandeis University. <http://www.brandeis.edu/wanghlab/>.
Hartshorn, C., Anshelevich, A., and Wangh, L.J.. "Laser zona-drilling does not induce hsp70i transcription in blastomeres of 8-cell mouse embryos." Fertility and Sterility 84. (5) (2005): 1547-1550.
Hartshorn, C., Anshelevich, A., and Wangh, L.J.. "Rapid, single-tube method for quantitative preparation and analysis of RNA and DNA in samples as small as one cell." BMC Biotechnology 5. (2005): 2.
Pierce, K.E., Sanchez, J.A., Rice, J.E., and Wangh, L.J. "Linear-After-The-Exponential (LATE)-PCR: Primer design criteria for high yields of specific single-stranded DNA and improved real-time detection." PNAS 102:. (2005): 8609-8614.
Pierce, K.E., and Wangh, L.J.. "Effectiveness and limitations of uracil-DNA glycosylases in sensitive real-time PCR assays." BioTechniques 36. (1) (2004): 44-48.
Sanchez, J.A., Pierce, K.E., Rice, J.E., and Wangh, L.J. "Linear-After-The-Exponential (LATE)-PCR: An advanced method of asymmetric PCR and its uses in quantitative real-time analysis." PNAS 101. (7) (2004): 1933-1938.
Hartshorn C, Rice JE, Wangh LJ.. "Differential pattern of Xist RNA accumulation in single blastomeres isolated from 8-cell stage mouse embryos following laser zona drilling." Molecular Reproduction and Development 64. (2003): 41-51.
Pierce, K.E., Rice, J.E., Sanchez, J.A., and Wangh, L.J.. "Detection of cystic fibrosis alleles from single cells using molecular beacons and a novel method of asymmetric real-time PCR." Molecular Human Reproduction 9. (12) (2003): 815-820.
Hartshorn, C., Rice, J.E., and Wangh, L.J.. "Developmentally-Regulated Changes of Xist RNA Levels in Single Preimplantation Mouse Embryos, as Revealed by Quantitative Real-Time PCR." Molecular Reproduction and Development 61. (2002): 425-436.
Pierce, K., Rice, J.E., Sanchez, J.A., and Wangh, L.J.. "QuantiLyse: Reliable DNA amplification from single cells." BioTechniques 32. (5) (2002): 1106-1111.
Rice, J.E., Sanchez, J.A., Pierce, K.E., and Wangh, L.J.. "Real-time PCR with molecular beacons provides a highly accurate assay for detection of Tay-Sachs alleles in single cells." Prenatal Diagnosis 22. (2002): 113-1134.
Pierce, K.E., Rice, J.E., Sanchez, J.A., Brenner, C., and Wangh, L.J.. "Real-time PCR using molecular beacons for accurate detection of the Y chromosome in single human blastomeres." Molecular Human Genetics 6. (12) (2000): 1155-1164.