Listeriosis laboratory tests: Difference between revisions
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==Overview== | ==Overview== | ||
For symptomatic patients, diagnosis is confirmed | For symptomatic patients, diagnosis of listeriosis is confirmed following the isolation of ''[[Listeria monocytogenes]]'' from a normally [[sterile]] site, such as [[blood]], [[spinal fluid]] (in the setting of [[nervous system]] involvement), or [[amniotic fluid]]/[[placenta]] (in the setting of [[pregnancy]]). Cultures from non-sterile sites, such as [[stool]] samples, are not recommended (1-15% carriage rate) but may still be useful in [[gastroenteritis]] with high suspicion of listeriosis. ''[[Listeria monocytogenes]]'' may be isolated readily on routine media. Since ''[[Listeria]]'' is an [[intracellular]] [[organism]], only 1/3 of cultures yield positive Gram-stains. Selective enrichment media improve rates of isolation from contaminated specimens. The cultures typically require 1-2 days for growth. A negative culture does not rule out [[infection]] in the presence of strong clinical suspicion. [[CSF analysis|Cerebrospinal fluid (CSF) analysis]] may confirm the diagnosis among patients with [[CNS]] listeriosis. [[Serology|Serological tests]] (e.g. [[listeriolysin O]] titers) have been used, but their use remains controversial and are currently not recommended. [[Polymerase chain reaction]] for the detection of the ''HLY'' gene may be diagnostic, but it is not yet widely available for commercial use. Laboratory testing on asymptomatic patients (including high-risk asymptomatic patients) is not recommended.<ref name=CDC>{{cite web | title = Listeria | url = http://www.cdc.gov/listeria/diagnosis.html }}</ref> | ||
==Laboratory Tests== | |||
'''The gold standard for the diagnosis of listeriosis is culture from sterile sites.''' | |||
===Culture=== | |||
*Diagnosis of listeriosis is made by culturing ''[[Listeria monocytogenes|Listeria]]'' from [[sterile]] sites (e.g. [[blood]], [[Cerebrospinal fluid|spinal fluid]]). | |||
*Cultures from non-sterile sites, such as [[stool]] culture or [[vaginal]] culture, are not helpful for the diagnosis of listeriosis (approximately 5% to 15% fecal carriage, especially among patients who receive [[PPI]] therapy).<ref name="pmid6701102">{{cite journal| author=Lennon D, Lewis B, Mantell C, Becroft D, Dove B, Farmer K et al.| title=Epidemic perinatal listeriosis. | journal=Pediatr Infect Dis | year= 1984 | volume= 3 | issue= 1 | pages= 30-4 | pmid=6701102 | doi= | pmc= | url=http://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=6701102 }} </ref> | |||
*[[Gram stain|Gram-stain]] may yield positive results in approximately 1/3 of infected patients (''[[Listeria monocytogenes|Listeria]]'' is an [[intracellular]] organism). | |||
* ''[[Listeria monocytogenes|Listeria]]'' grows on media such as Mueller-Hinton agar. | |||
* Identification is enhanced if the primary cultures are performed on agar containing sheep blood given the characteristic small zone of [[hemolysis]] that can be observed around, and under the colonies. | |||
* Isolation can be enhanced if the tissue is kept at 4°C for some days before inoculation into bacteriologic media. | |||
* The [[motility]] at room temperature and [[hemolysin]] production are primary findings that help differentiate [[Listeria monocytogenes|''Listeria'']] from other organisms (e.g. coryneform bacteria). | |||
*The cultures typically require 1-2 days for growth. | |||
===Stool Cultures=== | |||
*[[Stool]] cultures are not indicated in systemic [[listeriosis]] patients because routine culture media for enteric [[pathogens]] are not appropriate for the growth of ''[[Listeria monocytogenes|Listeria]]''. | |||
*In cases of outbreaks of [[listeriosis]] or individual patients with suspected listerial [[gastroenteritis]], special selected media can be used. | |||
===Listeriolysin O Titers=== | |||
*Elevated titers of [[listeriolysin O]] may distinguish patients with active ''[[Listeria monocytogenes|Listeria]]'' infections from those who are carriers of the [[organism]]. | |||
*The use of [[listeriolysin O]] for the diagnosis of listeriosis is still controversial.<ref name="pmid8972666">{{cite journal| author=Salamina G, Dalle Donne E, Niccolini A, Poda G, Cesaroni D, Bucci M et al.| title=A foodborne outbreak of gastroenteritis involving Listeria monocytogenes. | journal=Epidemiol Infect | year= 1996 | volume= 117 | issue= 3 | pages= 429-36 | pmid=8972666 | doi= | pmc=PMC2271639 | url=http://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=8972666 }} </ref><ref name="pmid8988887">{{cite journal| author=Dalton CB, Austin CC, Sobel J, Hayes PS, Bibb WF, Graves LM et al.| title=An outbreak of gastroenteritis and fever due to Listeria monocytogenes in milk. | journal=N Engl J Med | year= 1997 | volume= 336 | issue= 2 | pages= 100-5 | pmid=8988887 | doi=10.1056/NEJM199701093360204 | pmc= | url=http://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=8988887 }} </ref> | |||
===CSF Analysis=== | |||
*[[CSF]] analysis may confirm the diagnosis of [[listeriosis]]. Common findings include: | |||
:* [[Pleocytosis]] | |||
:* More than 25 [[lymphocytes]] in [[CSF]] [[Differential blood count (patient information)|differential count]], without [[antibiotic]] therapy | |||
:* Moderately elevated [[CSF]] [[protein]] concentration with reduced [[CSF]] [[glucose]] concentration<ref name="pmid9772921">{{cite journal| author=Mylonakis E, Hohmann EL, Calderwood SB| title=Central nervous system infection with Listeria monocytogenes. 33 years' experience at a general hospital and review of 776 episodes from the literature. | journal=Medicine (Baltimore) | year= 1998 | volume= 77 | issue= 5 | pages= 313-36 | pmid=9772921 | doi= | pmc= | url=http://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=9772921 }} </ref> | |||
* Normally, [[CSF analysis]] in the case [[bacterial meningitis]] reveals about 80% [[neutrophils]]. However, in [[Listeria]] [[meningitis]], [[neutrophils]] make up <80% of the [[White blood cell (WBC) count|WBCs]].<ref name="pmid8592552">{{cite journal |vauthors=Southwick FS, Purich DL |title=Intracellular pathogenesis of listeriosis |journal=N. Engl. J. Med. |volume=334 |issue=12 |pages=770–6 |year=1996 |pmid=8592552 |doi=10.1056/NEJM199603213341206 |url=}}</ref><ref name="pmid8416268">{{cite journal |vauthors=Durand ML, Calderwood SB, Weber DJ, Miller SI, Southwick FS, Caviness VS, Swartz MN |title=Acute bacterial meningitis in adults. A review of 493 episodes |journal=N. Engl. J. Med. |volume=328 |issue=1 |pages=21–8 |year=1993 |pmid=8416268 |doi=10.1056/NEJM199301073280104 |url=}}</ref><ref name="pmid3105048">{{cite journal |vauthors=Hansen PB, Jensen TH, Lykkegaard S, Kristensen HS |title=Listeria monocytogenes meningitis in adults. Sixteen consecutive cases 1973-1982 |journal=Scand. J. Infect. Dis. |volume=19 |issue=1 |pages=55–60 |year=1987 |pmid=3105048 |doi= |url=}}</ref> | |||
* In [[Listeria]] [[meningoencephalitis]], [[CSF analysis]] reveals 80-90% [[monocytes]].<ref name="pmid8592552">{{cite journal |vauthors=Southwick FS, Purich DL |title=Intracellular pathogenesis of listeriosis |journal=N. Engl. J. Med. |volume=334 |issue=12 |pages=770–6 |year=1996 |pmid=8592552 |doi=10.1056/NEJM199603213341206 |url=}}</ref><ref name="pmid8496712">{{cite journal |vauthors=Uldry PA, Kuntzer T, Bogousslavsky J, Regli F, Miklossy J, Bille J, Francioli P, Janzer R |title=Early symptoms and outcome of Listeria monocytogenes rhombencephalitis: 14 adult cases |journal=J. Neurol. |volume=240 |issue=4 |pages=235–42 |year=1993 |pmid=8496712 |doi= |url=}}</ref> | |||
* [[Gram stain]] of the [[CSF]] has very low [[sensitivity]]. Even when organisms are observed, they may be misidentified.<ref name="pmid4998254">{{cite journal| author=Lavetter A, Leedom JM, Mathies AW, Ivler D, Wehrle PF| title=Meningitis due to Listeria monocytogenes. A review of 25 cases. | journal=N Engl J Med | year= 1971 | volume= 285 | issue= 11 | pages= 598-603 | pmid=4998254 | doi=10.1056/NEJM197109092851103 | pmc= | url=http://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=4998254 }} </ref> Therefore, ''[[Listeria monocytogenes]]'' should always be considered when similar [[organisms]] are growing in [[blood culture |blood]] or [[CSF]] cultures. The presumptive diagnosis of [[viral meningitis]] should also be carefully considered in [[immunocompromised]], chronically ill, or elderly patients, presenting with acute [[meningitis]] and a negative [[Gram stain]]. In rhombencephalitis patients diagnosis is often delayed because [[CSF]] examination reveals only mild changes. | |||
===Polymerase Chain Reaction (PCR)=== | |||
* In [[CSF]] samples, [[polymerase chain reaction]] assay has been developed for the ''HLY'' gene detection, which encodes for the [[listeriolysin O]]. | |||
* However, despite a high [[specificity]] and more [[sensitivity]], [[Polymerase chain reaction|PCR]] for the diagnosis of listeriosis is not yet widely available for commercial use. | |||
==References== | ==References== | ||
{{reflist|2}} | {{reflist|2}} | ||
{{ | {{WH}} | ||
{{ | {{WS}} | ||
[[Category: | [[Category:Emergency mdicine]] | ||
[[Category:Disease]] | [[Category:Disease]] | ||
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[[Category:Infectious disease]] | [[Category:Infectious disease]] | ||
[[Category:Neurology]] | |||
[[Category:Gastroenterology]] |
Latest revision as of 22:31, 29 July 2020
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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1]; Associate Editor(s)-in-Chief: João André Alves Silva, M.D. [2]
Overview
For symptomatic patients, diagnosis of listeriosis is confirmed following the isolation of Listeria monocytogenes from a normally sterile site, such as blood, spinal fluid (in the setting of nervous system involvement), or amniotic fluid/placenta (in the setting of pregnancy). Cultures from non-sterile sites, such as stool samples, are not recommended (1-15% carriage rate) but may still be useful in gastroenteritis with high suspicion of listeriosis. Listeria monocytogenes may be isolated readily on routine media. Since Listeria is an intracellular organism, only 1/3 of cultures yield positive Gram-stains. Selective enrichment media improve rates of isolation from contaminated specimens. The cultures typically require 1-2 days for growth. A negative culture does not rule out infection in the presence of strong clinical suspicion. Cerebrospinal fluid (CSF) analysis may confirm the diagnosis among patients with CNS listeriosis. Serological tests (e.g. listeriolysin O titers) have been used, but their use remains controversial and are currently not recommended. Polymerase chain reaction for the detection of the HLY gene may be diagnostic, but it is not yet widely available for commercial use. Laboratory testing on asymptomatic patients (including high-risk asymptomatic patients) is not recommended.[1]
Laboratory Tests
The gold standard for the diagnosis of listeriosis is culture from sterile sites.
Culture
- Diagnosis of listeriosis is made by culturing Listeria from sterile sites (e.g. blood, spinal fluid).
- Cultures from non-sterile sites, such as stool culture or vaginal culture, are not helpful for the diagnosis of listeriosis (approximately 5% to 15% fecal carriage, especially among patients who receive PPI therapy).[2]
- Gram-stain may yield positive results in approximately 1/3 of infected patients (Listeria is an intracellular organism).
- Listeria grows on media such as Mueller-Hinton agar.
- Identification is enhanced if the primary cultures are performed on agar containing sheep blood given the characteristic small zone of hemolysis that can be observed around, and under the colonies.
- Isolation can be enhanced if the tissue is kept at 4°C for some days before inoculation into bacteriologic media.
- The motility at room temperature and hemolysin production are primary findings that help differentiate Listeria from other organisms (e.g. coryneform bacteria).
- The cultures typically require 1-2 days for growth.
Stool Cultures
- Stool cultures are not indicated in systemic listeriosis patients because routine culture media for enteric pathogens are not appropriate for the growth of Listeria.
- In cases of outbreaks of listeriosis or individual patients with suspected listerial gastroenteritis, special selected media can be used.
Listeriolysin O Titers
- Elevated titers of listeriolysin O may distinguish patients with active Listeria infections from those who are carriers of the organism.
- The use of listeriolysin O for the diagnosis of listeriosis is still controversial.[3][4]
CSF Analysis
- CSF analysis may confirm the diagnosis of listeriosis. Common findings include:
- Pleocytosis
- More than 25 lymphocytes in CSF differential count, without antibiotic therapy
- Moderately elevated CSF protein concentration with reduced CSF glucose concentration[5]
- Normally, CSF analysis in the case bacterial meningitis reveals about 80% neutrophils. However, in Listeria meningitis, neutrophils make up <80% of the WBCs.[6][7][8]
- In Listeria meningoencephalitis, CSF analysis reveals 80-90% monocytes.[6][9]
- Gram stain of the CSF has very low sensitivity. Even when organisms are observed, they may be misidentified.[10] Therefore, Listeria monocytogenes should always be considered when similar organisms are growing in blood or CSF cultures. The presumptive diagnosis of viral meningitis should also be carefully considered in immunocompromised, chronically ill, or elderly patients, presenting with acute meningitis and a negative Gram stain. In rhombencephalitis patients diagnosis is often delayed because CSF examination reveals only mild changes.
Polymerase Chain Reaction (PCR)
- In CSF samples, polymerase chain reaction assay has been developed for the HLY gene detection, which encodes for the listeriolysin O.
- However, despite a high specificity and more sensitivity, PCR for the diagnosis of listeriosis is not yet widely available for commercial use.
References
- ↑ "Listeria".
- ↑ Lennon D, Lewis B, Mantell C, Becroft D, Dove B, Farmer K; et al. (1984). "Epidemic perinatal listeriosis". Pediatr Infect Dis. 3 (1): 30–4. PMID 6701102.
- ↑ Salamina G, Dalle Donne E, Niccolini A, Poda G, Cesaroni D, Bucci M; et al. (1996). "A foodborne outbreak of gastroenteritis involving Listeria monocytogenes". Epidemiol Infect. 117 (3): 429–36. PMC 2271639. PMID 8972666.
- ↑ Dalton CB, Austin CC, Sobel J, Hayes PS, Bibb WF, Graves LM; et al. (1997). "An outbreak of gastroenteritis and fever due to Listeria monocytogenes in milk". N Engl J Med. 336 (2): 100–5. doi:10.1056/NEJM199701093360204. PMID 8988887.
- ↑ Mylonakis E, Hohmann EL, Calderwood SB (1998). "Central nervous system infection with Listeria monocytogenes. 33 years' experience at a general hospital and review of 776 episodes from the literature". Medicine (Baltimore). 77 (5): 313–36. PMID 9772921.
- ↑ 6.0 6.1 Southwick FS, Purich DL (1996). "Intracellular pathogenesis of listeriosis". N. Engl. J. Med. 334 (12): 770–6. doi:10.1056/NEJM199603213341206. PMID 8592552.
- ↑ Durand ML, Calderwood SB, Weber DJ, Miller SI, Southwick FS, Caviness VS, Swartz MN (1993). "Acute bacterial meningitis in adults. A review of 493 episodes". N. Engl. J. Med. 328 (1): 21–8. doi:10.1056/NEJM199301073280104. PMID 8416268.
- ↑ Hansen PB, Jensen TH, Lykkegaard S, Kristensen HS (1987). "Listeria monocytogenes meningitis in adults. Sixteen consecutive cases 1973-1982". Scand. J. Infect. Dis. 19 (1): 55–60. PMID 3105048.
- ↑ Uldry PA, Kuntzer T, Bogousslavsky J, Regli F, Miklossy J, Bille J, Francioli P, Janzer R (1993). "Early symptoms and outcome of Listeria monocytogenes rhombencephalitis: 14 adult cases". J. Neurol. 240 (4): 235–42. PMID 8496712.
- ↑ Lavetter A, Leedom JM, Mathies AW, Ivler D, Wehrle PF (1971). "Meningitis due to Listeria monocytogenes. A review of 25 cases". N Engl J Med. 285 (11): 598–603. doi:10.1056/NEJM197109092851103. PMID 4998254.