Thanks for the clarification, Jimmy!


*From:* [email protected] <[email protected]>
*On Behalf Of *Jimmy Eng
*Sent:* Friday, December 13, 2019 8:27 AM
*To:* [email protected]
*Subject:* Re: [spctools-discuss] Interpretation of stats from pepXML viewer



Regarding inputs and output counts to Comet, as Eric mentioned, there are
search parameters which affect whether or not a spectrum will be searched
at all.  These are: activation_method, digest_mass_range,
max_precursor_charge, minimum_peaks, minimum_intensity, and
remove_precursor_peak (which can affect minimum_peaks).  A query spectrum
that happens to be out of bounds for any of these parameters will not be
analyzed (is effectively ignored) and will not show up in the output.  Any
spectra that passes these filters will have an output entry in Comet's
pepXML output file, even if there's no matching peptide output.  Such blank
outputs can happen in cases where the database or tolerances are so small
that no peptides are scored or the spectrum is so poor that the xcorr score
is less than or equal to zero.  This is why you see 1752 spectra in the
pepXML after Comet searches of both your databases as that's the number of
spectra that actually get analyzed and is a function of the input and not
the database being searched.



On Fri, Dec 13, 2019 at 2:29 AM 'Alastair Skeffington' via spctools-discuss
<[email protected]> wrote:

Hi,



Can someone explain what's happen here please:



Number of spectra in the .mgf file = 1856

Number of spectra after conversion to mzXML = 1856

Number of spectra in pepXML file after comet search: 1752



I then search against database A (DB-A) and database B (DB-B) and count the
number of spectra in the pepXML file as counts of "<spectrum_query
spectrum=". Yields 1752 in both cases.



I then run "xinteract -Neg.pepXML -PPM -dDECOY_ -OARP eg_pepp.pepXML" on
each of the files. Now when I search for the string "<spectrum_query
spectrum=" I get 481 hits for DB-A and 632 hits for DB-B. The pepXML view
then calculates the "Efficiency ID'd/searched" - so the denominator is
different for each database.



Can someone help to explain this behavior?



   - Why does the xinteract output present different numbers of records
   ("<spectrum_query spectrum= ...") for the two different databases, even
   though the number of records in the pepXML input was the same?
   - Isn't the definition of 'Spectra searched' used by Petunia misleading?
   It's not the number of input spectra to the search engine.
   - Presumably the loss of spectra between the input to comet and the
   output is due to loosing those spectra with absolutely no match to the
   database or decoys due to some internal threshold on the score for PSMs.



Any help would be hugely appreciated!



Thanks,

Alastair

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