Monkeypox laboratory tests: Difference between revisions

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(Created page with "__NOTOC__ {{Monkeypox}} {{CMG}} {{AE}} Bassel Almarie, M.D. ==Overview== The laboratory findings of chickenpox include blood tests that can be done to identify the response to acute infection (IgM) or previous infection and subsequent immunity (IgG). Prenatal diagnosis of fetal varicella infection can be performed using ultrasound at 5 weeks following primary maternal infection. A ...")
 
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==Laboratory Findings==
==Laboratory Findings==


Laboratory tests that are shown to be useful for the diagnosis of [[chickenpox]] include:
===Molecular Methods===
* [[Polymerase chain reaction|PCR]] is done by collecting [[skin]] [[Vesicle|vesicles]] [[fluid]], [[Scab|scabs]], [[saliva]] and [[cerebrospinal fluid]] if neurological symptoms or signs are present.<ref name="pmid27188665">{{cite journal| author=Gershon AA, Breuer J, Cohen JI, Cohrs RJ, Gershon MD, Gilden D et al.| title=Varicella zoster virus infection. | journal=Nat Rev Dis Primers | year= 2015 | volume= 1 | issue=  | pages= 15016 | pmid=27188665 | doi=10.1038/nrdp.2015.16 | pmc=5381807 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=27188665  }} </ref>
====Polymerase chain reaction (PCR)====
*[[Polymerase chain reaction|PCR]] done with a [[restriction enzyme]] can be used to [[hydrolyze]] and sequence specific segments of the [[Virus|viral]] [[genome]]. This can be used to determine whether [[Varicella zoster virus|VZV]] is resistant to [[acyclovir]].<ref name="pmid27188665">{{cite journal| author=Gershon AA, Breuer J, Cohen JI, Cohrs RJ, Gershon MD, Gilden D et al.| title=Varicella zoster virus infection. | journal=Nat Rev Dis Primers | year= 2015 | volume= 1 | issue=  | pages= 15016 | pmid=27188665 | doi=10.1038/nrdp.2015.16 | pmc=5381807 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=27188665  }} </ref>
* [[Antibody]] Testing
* [[Virus|Viral]] detection
* [[Varicella zoster virus|VZV]] [[DNA]] testing
* Direct [[Fluorescent]] Antibody ([[Direct fluorescent antibody|DFA]])
* [[Varicella zoster virus|VZV]] culture


===Antibody Testing===
*Confirmation of monkeypox virus (MPXV) infection is based on nucleic acid amplification testing, using real-time or conventional PCR, for detection of unique sequences of viral DNA. PCR can be used alone, or in combination with sequencing.
*Laboratory tests detect and measure the level of [[Varicella zoster virus|VZV]] [[antibodies]]. The presence of [[IgM]] antibodies indicates acute [[infection]].
*PCR kits detecting MPXV are under development but no commercial validated PCR kits are currently available widely.


====IgM====
====DNA extraction ====
*It can be detected within 1-2 weeks post exposure.
 
*The levels of [[Immunoglobulin M|IgM antibodies]] rise for a short period of time and then fall below detectable levels.
*DNA can be extracted from samples using any standard extraction protocols or kits. In general, the sample lysis step in DNA extraction inactivates any live virus. Thus, it is recommended that the sample lysis step is performed under a biosafety cabinet. For crust samples, DNA extraction kit for tissue samples should be used to insure appropriate sample lysis.
*Post-Infection, the [[IgM]] levels rise only when the latent [[VZV]] is reactivated.
====IgG====
*[[Immunoglobulin G|IgG antibodies]] are produced several weeks after the initial exposure.
*[[IgG]] levels rise during active [[infection]] and then the levels become stable as the [[Varicella zoster virus|VZV]] infection gets resolves and as the [[virus]] gets inactivated.
*[[IgG]] levels rise during active [[infection]] and then the levels become stable as the [[Varicella zoster virus|VZV]] infection gets resolves and as the [[virus]] gets inactivated.


===Viral detection===
===Viral detection===
* [[Virus|Viral]] detection is done by finding [[Varicella zoster virus|VZV]] in a [[blood]], [[vesicle]] [[fluid]], or [[tissue]] sample. Detection is done by culturing the [[virus]] or by detecting its [[genetic material]] ([[Varicella zoster virus|VZV]] [[DNA]]).
 
*[[Virus|Viral]] detection is done by finding [[Varicella zoster virus|VZV]] in a [[blood]], [[vesicle]] [[fluid]], or [[tissue]] sample. Detection is done by culturing the [[virus]] or by detecting its [[genetic material]] ([[Varicella zoster virus|VZV]] [[DNA]]).


===VZV DNA testing===
===VZV DNA testing===


* [[Varicella zoster virus|VZV]] [[DNA]] testing is [[Sensitivity (tests)|sensitive]] and measures [[viral load]].
*[[Varicella zoster virus|VZV]] [[DNA]] testing is [[Sensitivity (tests)|sensitive]] and measures [[viral load]].


===Direct Fluorescent Antibody (DFA)===
===Direct Fluorescent Antibody (DFA)===


* [[Direct fluorescent antibody|DFA]] test detects the presence of [[Varicella zoster virus|VZV]] in the cells in the [[skin]] lesion using a labeled [[antibody]]. It is rapid but less [[Specificity (tests)|specific]] and [[Sensitivity (tests)|sensitive]] than [[DNA]] testing.
*[[Direct fluorescent antibody|DFA]] test detects the presence of [[Varicella zoster virus|VZV]] in the cells in the [[skin]] lesion using a labeled [[antibody]]. It is rapid but less [[Specificity (tests)|specific]] and [[Sensitivity (tests)|sensitive]] than [[DNA]] testing.


===VZV culture===
===VZV culture===


* [[Culture medium|Culture]] is not very reliable for [[Varicella zoster virus|VZV]] and can lead to [[false-negative]] results.
*[[Culture medium|Culture]] is not very reliable for [[Varicella zoster virus|VZV]] and can lead to [[false-negative]] results.


===Microscopic Findings===
===Microscopic Findings===


<gallery>
<gallery>
 
File:Chickenpox37.jpeg| Photomicrograph reveals the intranuclear inclusions produced by varicella virus grown in a tissue culture (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
Image: Chickenpox37.jpeg| Photomicrograph reveals the intranuclear inclusions produced by varicella virus grown in a tissue culture (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
File:Chickenpox14.jpeg|Cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox (125x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
File:Chickenpox13.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (125x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
Image: Chickenpox14.jpeg|Cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox (125x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
File:Chickenpox12.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
File:Chickenpox11.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
Image: Chickenpox13.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (125x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
File:Chickenpox10.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
File:Chickenpox09.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
Image: Chickenpox12.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
File:Chickenpox08.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
File:Chickenpox07.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name="PHIL"> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
Image: Chickenpox11.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
Image: Chickenpox10.jpeg|Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
Image: Chickenpox09.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
Image: Chickenpox08.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
Image: Chickenpox07.jpeg|Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). <SMALL><SMALL>''[http://phil.cdc.gov/phil/home.asp From Public Health Image Library (PHIL).] ''<ref name=PHIL> {{Cite web | title = Public Health Image Library (PHIL) | url = http://phil.cdc.gov/phil/home.asp}}</ref></SMALL></SMALL>
 
</gallery>
</gallery>


Revision as of 15:17, 15 June 2022

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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1] Associate Editor(s)-in-Chief: Bassel Almarie, M.D.

Overview

The laboratory findings of chickenpox include blood tests that can be done to identify the response to acute infection (IgM) or previous infection and subsequent immunity (IgG). Prenatal diagnosis of fetal varicella infection can be performed using ultrasound at 5 weeks following primary maternal infection. A PCR test of the mother's amniotic fluid can also be performed, though the risk of spontaneous abortion due to the amniocentesis procedure is higher than the risk of the baby developing fetal varicella syndrome.

Laboratory Findings

Molecular Methods

Polymerase chain reaction (PCR)

  • Confirmation of monkeypox virus (MPXV) infection is based on nucleic acid amplification testing, using real-time or conventional PCR, for detection of unique sequences of viral DNA. PCR can be used alone, or in combination with sequencing.
  • PCR kits detecting MPXV are under development but no commercial validated PCR kits are currently available widely.

DNA extraction

  • DNA can be extracted from samples using any standard extraction protocols or kits. In general, the sample lysis step in DNA extraction inactivates any live virus. Thus, it is recommended that the sample lysis step is performed under a biosafety cabinet. For crust samples, DNA extraction kit for tissue samples should be used to insure appropriate sample lysis.
  • IgG levels rise during active infection and then the levels become stable as the VZV infection gets resolves and as the virus gets inactivated.

Viral detection

VZV DNA testing

Direct Fluorescent Antibody (DFA)

VZV culture

Microscopic Findings

  • Photomicrograph reveals the intranuclear inclusions produced by varicella virus grown in a tissue culture (500x mag). From Public Health Image Library (PHIL). [1]

    Photomicrograph reveals the intranuclear inclusions produced by varicella virus grown in a tissue culture (500x mag). From Public Health Image Library (PHIL). [1]

  • Cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox (125x mag). From Public Health Image Library (PHIL). [1]

    Cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox (125x mag). From Public Health Image Library (PHIL). [1]

  • Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (125x mag). From Public Health Image Library (PHIL). [1]

    Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (125x mag). From Public Health Image Library (PHIL). [1]

  • Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). From Public Health Image Library (PHIL). [1]

    Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). From Public Health Image Library (PHIL). [1]

  • Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). From Public Health Image Library (PHIL). [1]

    Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). From Public Health Image Library (PHIL). [1]

  • Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). From Public Health Image Library (PHIL). [1]

    Hematoxylin-eosin (H&E)-stained photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (50x mag). From Public Health Image Library (PHIL). [1]

  • Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). From Public Health Image Library (PHIL). [1]

    Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (500x mag). From Public Health Image Library (PHIL). [1]

  • Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). From Public Health Image Library (PHIL). [1]

    Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). From Public Health Image Library (PHIL). [1]

  • Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). From Public Health Image Library (PHIL). [1]

    Photomicrograph reveals some of the cytoarchitectural histopathologic changes which you’d find in a human skin tissue specimen that included a chickenpox, or varicella zoster virus lesion (1200x mag). From Public Health Image Library (PHIL). [1]

References

  1. ↑ 1.0 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 "Public Health Image Library (PHIL)".

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