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Research Materials Available from FR3 Molecular Resources

Filarial RNA and DNA preparations

TO ORDER FILARIAL RNA AND DNA PREPARATIONS, YOU MUST HAVE A CURRENT FR3 REGISTRATION FORM ON FILE, AND A SMITH COLLEGE MTA.  Currently, RNA and DNA preparations are ordered directly through Smith College (genome@smith.edu).

Species Preparation  Life Cycle Stage(s)
Brugia malayi total RNA  adult female
adult male
L3
L4
microfilaria 
B. pahangi total RNA adult male
microfilaria
Dirofilaria immitis total RNA adult female
L3 
Aedes aegypti total RNA eggs
male pupae
young adult
adult male
adult female
B. malayi DNA  ----- 
D. immitis DNA  ----- 
Acanthocheilonema viteae DNA  ----- 
Aedes aegypti DNA  ----- 
Wuchereria bancrofti DNA  whole genome amplified from mf 

Please contact  (genome@smith.edu) to order these new reagents!!!

 

cDNA LIBRARIES - cDNA CLONES FROM INDICATED (*) LIBRARIES MUST BE ORDERED FROM BEI LABORATORIES (www.beiresources.org).

Entire libraries can still be ordered through FR3.  Fill out the BEI Resources Material Transfer Agreement and Registration, then use the 'FR3 Ordering' link on the BEI homepage to get started.

Characteristics of the Onchocerca volvulus Microfilaria cDNA Library.  Please click each library to enter its data sheet.
 
Onchocerca volvulus  Microfilarial cDNA Library (SAW98MLW-OvMf)*
 L2 cDNA Library (SAW98MLW-OvL2)*
 Infective L3 cDNA Library (SAW94WL-OvL3)*
 Adult female cDNA Library (SAW98MLW-OvAF)*
 Adult male cDNA Library (SAW98MLW-OvAM)*
 Adult female ivermectin treated cDNA Library (SAW99PF-OvAF)
 Genomic library (OvG)
Molting L3 cDNA Library (Ov3M)*
Onchocerca ochengi  Infective L3 cDNA Library (SAW98MLW-OoL3)
Brugia malayi  Microfilarial cDNA library (SAW94LS-BmMf)
 L2 cDNA library (JHU96SL-BmL2)
 Infective L3 cDNA library (SAW94WL-BmL3)
 L3 molting-day 6 larval cDNA library  (SAW96MLW-BmL3d6)
 L3 molting-day 9 larval cDNA library (SAW96MLW-BmL3d9)
 L3 subtracted cDNA library (SAW97LS-BmL3SA)
 L3 subtracted cDNA library (SAW97YG-BmL3SB)*
 L3 subtracted cDNA library (SAW97LS-BmL3SD)*
 L4 cDNA library (SAW99MLW--BmL4)*
 Young adult cDNA library (SAW99MLW-BmYA)*
 Adult male cDNA library (SAW94NL-BmAM)*
 Adult female cDNA library (SAW96MLW-BmAF)*
 Adult cDNA library (JHU96SL-BmA)
 Genomic library (BmG)
Wuchereria bancrofti  Microfilarial cDNA Library (SAW95SJL-WbMf)*
 Infective L3 cDNA Library (SAW96MLW-WbL3)
 Adult female cDNA Library (JHU94SL-fWb)
 Adult male cDNA Library (JHU94SL-mWb)

 
Library Name   SAW98MLW-OvMf
 Key: SAW Lab:
SAW = Steven A. Williams' Lab
JHU = Johns Hopkins University - Alan Scott's Lab
  98 year constructed
  MLW initials of the individual who constructed the library
  Ov species:
Ov = Onchocerca volvulus
Oo = Onchocerca ochengi
Bm = Brugia malayi
Wb = Wuchereria bancrofti
  Mf stage:
Mf = microfilaria
L2 = second larval stage
L3 = third larval stage
L3d6 = L3 to L4 molt, day 6
L3d9 = L3 to L4 molt, day 9
L3SA (L3SD) = L3 cDNA subtracted with microfilaria, adult male and
adult female cDNA
L3SB = L3 cDNA obtained by subtracting JHU93SL-BmL3 cDNA library (constructed using SL sequence and PCR) from SAW94WL-BmL3 cDNA library (constructed using conventional cDNA methods).
L4 = fourth larval stage
YA = young adult
AF = adult female
AM = adult male

 

 

B. malayi MICROARRAY

TO ORDER THE BRUGIA MICROARRAY, YOU MUST HAVE A CURRENT FR3 REGISTRATION FORM ON FILE, AND A SMITH COLLEGE MTA. 

The second version (V.2) of the filarial microarrays are now available through the Molecular Division of FR3.  These can be requested just like any other reagent using the Requisition Form found on this site.   The only additional requirement is that the individual lab requesting the arrays must summarize briefly the experiment for which the arrays are being requested. Please see the MIAME protocol and checklist.   This is so I can inform the lab requesting the arrays if another lab is already doing a substantially similar experiment.   This will not prevent any lab from doing any experiment they wish, it is merely to keep everyone informed of any duplication of effort.   This may enable laboratories to modify their plans in order to ask slightly different questions.

The arrays are made available through the efforts and funding of the Filarial Microarray Consortium (FMC).   The FMC includes the following individuals who contributed funds for the design and printing of the arrays: Steven A. Williams (Smith College), Gary Weil (Washington University), David Spiro (TIGR), Barton Slatko (New England Biolabs), Thomas Nutman (NIAID), Thomas Klei (LSU), Achim Hoerauf (University of Bonn), Sara Lustigman (New York Blood Center), Brenda Beerntsen (U. Missouri), TV Rajan (U. Connecticut Health Center), and Alan Scott (Johns Hopkins). Special thanks to David Spiro and Seth Crosby ( Washington University) for their hard work in much of the design of the arrays.

The 18,153 Oligonucleotides on the Version 2 (V.2) array include:

  • 9,241 oligos based on the TIGR B. malayi gene models (predicted genes)
  • 1,889 oligos based on TIGR gene indices (EST clusters and singletons not represented in the gene models above)
  • 81 oligos based on NEMBASE EST clusters (those not already represented above)
  • 675 oligos based on EST clusters from L3 EST clusters from Wash. U. Parasite Genomes (those not already represented above)
  • 878 oligos based on EST clustering of W. bancrofti data (those not already represented above)
  • 1,016 oligos based TIGR O. volvulus gene indices (EST clusters and singletons not already represented above)
  • 804 oligos based on the search of the Wolbachia complete genome for open reading frames
  • 3,569 oligos based on the original B. malayi EST clusters from the version 1 array (McPherson oligos)

The B. malayi microarray platform has been extensively validated by quantitative reverse transcriptase polymerase chain reaction (Li et al. 2005Mol. Biochem. Parasitol. 143:49-57) and tissue localization by in situ hybridization (Jiang et al. 2008Int. J. Parasitol. 38:503-12). For further information, consult the Filarial Genomics and Bioinformatics page, or contact Steven A. Williams, Director, Molecular Division FR3 at genome@smith.edu.

 

GENOMIC LIBRARIES

B. malayi BAC library clones - used to map the Wolbachia genome (see Foster et al. 2004. Int. J. Parasit. 34: 733-746; and Foster et al. 2001BioTechniques 30(6): 1216-1222).

 

FILARIAL SERUM SAMPLES/ANTIGENS

TO ORDER FILARIAL SERUM SAMPLES AND ANTIGENS, YOU MUST HAVE A CURRENT FR3 REGISTRATION FORM ON FILE, AND A SMITH COLLEGE MTA. 

Bm14 Antigen for serologic monitoring

Positive Control for the Binax Filariasis Now test

Filarial Serum Bank

 

BmWSP MONOCLONAL ANTIBODY - THIS MONOCLONAL IS AVAILABLE THROUGH BEI LABORATORIES (www.beiresources.org). 

Fill out the BEI Resources Material Transfer Agreement and Registration, then use the FR3 Ordering link on the BEI homepage to get started.  This monoclonal was developed by Patrick Lammie at Centers for Disease Control, and recognizes wBm0432.

Real-time PCR Assay for the Simultaneous Detection of Wuchereria bancrofti and Brugia malayi

 When conducted in accordance with the published protocol [1], this multiplex real-time PCR assay is capable of detecting Wuchereria bancrofti and Brugia malayi DNA simultaneously, or independently, at sensitivities which parallel the published singleplex assays for parasite detection.  This assay has been tested on worm extracted DNA, as well as DNA extracted from both human bloodspots and vector mosquito pools.  Xenomonitoring studies conducted in co-endemic locations can benefit from the reagent-cost and labor-cost related savings that this assay is able to provide.

1).  Pilotte N, Torres M, Tomaino FR, Laney SJ, Williams SA.  A TaqMan-based multiplex real-time PCR assay for the simultaneous detection of Wuchereria bancrofti and Brugia malayi.  Mol Biochem Parasitol 2013 May; 189(1-2): 33-37. 

Smith College will provide primers, controls and protocol for this procedure.  

Please contact  (genome@smith.edu) for further information regarding this assay.  

 

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